14 - 1 Cultivo Cel Animales Gibco Catalogue Edition 49

gibco cell culture catalogue
Introduction
When reliable results and confidence in your work are critical to your success, trust gibco products and services. Quality is built into every gibco product, from the finest raw materials to precise formulations and rigorous final product testing. Gibco products are consistent over time, lot to lot and bottle to bottle. To you, that means greater reliability, greater consistency, greater control, and ultimately, greater confidence in your results. Comprehensive range of products provides what you need, when you need it Lot to lot, bottle to bottle, gibco products are reliably consistent over time Gibco quality goes into every product, from the highest-quality raw materials to our rigorous final product testing Manufactured in cGMP facilities, certified to FDA Regulations 21 CFR Part 820 and ISO 13485:2003 to meet stringent quality standards New and innovative products designed to simplify your workflow and improve your Cells health and performance Advancing the robustness of your experiments through our leadership in enabling Advanced cell systems for research Our award-winning technical service group is your resource for products, techniques, troubleshooting and applications
Better results begin with better cell culture. To learn more about the gibco collection of Products and services, visit www.invitrogen.com/gibco
Resources web links

The gibcocell culture basics handbook: www.invitrogen.com/cellculturebasics cell culture protocol: www.invitrogen.com/protocols Information on mammalian and insect cell culture cell type specific protocols and additional cell culture products: www.invitrogen.com/cellculture Technical Support, Safety data sheets (SDS) are available at: www.invitrogen.com/sds Certificate of Analysis provides detailed quality control and products qualification information for each product. Certificates of Analysis are available on our website www.invitrogen.com/support, search for the certificate of analysis by product lot number which is printed on the box. Media Selection tools: http://igene.invitrogen.com/products/selector/pscs http://igene.invitrogen.com/products/selector/media
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Important Licensing Information: The products in this catalogue may covered by one or more Limited Use Label Licenses (see the Invitrogen website www.invitrogen.com). By use of these products, you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for research use only, and are not intended for human or animal diagnostics, therapeutic, commercial, or other clinical uses unless otherwise specifically indicated in the applicable product Limited Use Label License(s).
Introduction
Cell Culture Products

This information applies to the following cell culture product chapters, with the exception of Chapter 8, Animal Sera. The products listed in the following chapters are for research use and further cell culture manufacturing only, except where otherwise denoted. These products are not for diagnostic use. The safety and efficacy for these products in diagnostic or other clinical uses have not been established. In appropriate circumstances these products may be used as raw material components in further manufacturing applications. Products listed for in vitro diagnostic use are medical devices subject to the requirements of the Code of Federal Regulations Title 21, Part 820: Quality System Regulation. These products may also be used for research applications, and in appropriate circumstances, as raw material components in further manufacturing applications. It is the end users responsibility to qualify these products for their specific appli cation. They have not been qualified nor are they intended for any animal or human therapeutic uses. an AGT medium is complete and does not require pH/OSMO adjustment. Additionally, because the granules wet instantly, providing easier reconstitution, overall medium preparation time is reduced.
Containers and closures

The following flexible plastic bag packages are available for liquid media and most reagents: 1 L, 5 L, 10 L, 20 L, 100 L, and 200 L. Stock configurations of these units have been designed for general utility and convenience, but the customization of these bags to meet your specific needs is available. More information about our line of custom packaging can be obtained at www.invitrogn.com/synergy, through your Account Manager, or by contacting our Custom Products Services group.
Production and Preparation

Liquid cell culture products
Water preparation. The water used in liquid cell culture products is distilled water meeting all USP Monograph test requirements for Water For Injection. The system includes water pretreatment followed by distillation. The pure distillate is collected in storage tanks for holding until distribution via a constantly recirculating loop to points of use. The water is cooled to formulation temperature by a heat exchanger. All piping in the still, storage, and distribution systems is constructed of sanitary 316L stainless steel. The system is controlled automatically by a microprocess controller employing state-of-the-art sensing and fail-safe systems. Continuous quality assurance testing for microbial bioburden, endotoxin, and Water For Injection USP monograph requirements ensures that all specifications are met. Addition of chemicals. All raw material components used in our liquid medium formulations undergo stringent quality control tests, such as for identity and purity, to provide assurance that our media meet their final product quality control specifications. The biochemicals used in our formulations are USP, ACS, FCC, or other compendial grade. When no published standard is available, we set internal specifications (gibco grade) to ensure optimal product performance. Chemicals of confirmed high quality are weighed accurately and confirmed using a state-of-the-art computerized weighing system. The chemicals are then solubilized in distilled water in calibrated formulation tanks. In each batch preparation, mixing is maintained to ensure homogeneity. Membrane filtration. gibco cell culture media are prepared by an aseptic filling process for which each step has been validated to ensure the production of a product meeting the industry-standard sterility assurance level of 103 (i.e., product demonstrates a contamination level of no more than 1 of 1,000 units during the manufacturing process). The highest level of sterility assurance (equal to or greater than 106) cannot be achieved without terminal sterilization, which can be harmful to the performance of some cell culture products. Key points of control include validated sterilization cycles for all material with product contact, routine medium fills employing bacteriological media, a comprehensive environmental monitoring program, validated cleaning procedures, and a validated
Custom Cell Culture Products

We have the technical and manufacturing capabilities to produce a wide variety of customized cell culture media and reagents. From slight formulation modifications to complicated engineered-to-order products, custom products are available in both standard and highly specialized packaging configurations. Wherever possible, we can formulate custom media with animal originfree components and offer developmental support to help you re-engineer formulations to meet regulatory and performance goals. Gibco is the only brand that offers four distinct formats for media: ready-to-use (1X) liquid media, dry powder media (DPM), liquid medium concentrates (LMCs), and Advanced Granulation Technology (AGT). Whether you need media for cell culture or buffers for purification, submit your requests to our Custom Products Services group for timely evaluation and production of the formulation and delivery system that best fits your needs.
Custom liquid medium concentrates

Unique features of liquid medium concentrates include improved nutrient solubility, superior correlation with theoretical nutrient potency, and glutamine stabilization.
Advanced Granulation Technology (AGT)

AGT is a dry form of cell culture media in a new granular format that is optimally suited to research and industrial-scale production applications. By applying the well-characterized pharmaceutical manufacturing technology of fluid-bed granulation to cell culture media, we can now offer modern, highly complex formulations in an easy-to-use format. AGT media are complete and pH/OSMO adjusted. The essential benefit of a complete AGT medium is that it provides one raw material for the user and can decrease costs and time involved in raw material planning, procurement, and testing. Medium preparation is also simplified because
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final filter integrity testing program. Additionally, filtration and dispensing are performed within positive-pressure, HEPA-filtered, environmentally controlled rooms. The filled containers are labeled and maintained under quarantine at their required temperatures until quality control tests are complete and the product meets our final quality control release specifications. Prior to release, all production records and test results undergo a final review by our Quality Assurance Team.
Powdered Cell Culture Products

All raw material components used in our powdered medium formulations undergo stringent quality control tests, such as for identity and purity, to provide assurance that our media meet their final product quality control specifications. The biochemicals used in our formulations are USP, ACS, FCC, or other compendial grade. When no published standard is available we set internal specifications (gibco grade) to ensure optimal product performance.
inoculated at approximately 5 104 cells/ml and the logarithmic growth rate monitored relative to parallel cultures grown in a standardized reference medium. Specialized culture media are performance tested on appropriate cell lines compared to recognized reference standards. Endotoxin tests. Endotoxin values are measured with the Limulus Amebocyte Lysate (LAL) test. Media with the lowest level of endotoxin attainable in the industry today are routinely available. Stability testing program. The expiration date shown on every in vitro diagnostic (IVD)-labeled product is an indication of how long the product will deliver effective performance. Our stability program establishes and verifies product expiration dates. Product groups are monitored at periodic intervals to determine the length of time that the product will deliver acceptable performance under recommended storage conditions.
Powdered media
The quality of powdered media is confirmed by evaluating samples from each batch as required by specifications. Samples are prepared in 1X liquid form and membrane filtered, and testing may include: Growth promotion/toxicity Solubility Osmolality pH The values for these parameters are determined using the test conditions described for liquid media. All powdered media are produced without sodium bicarbonate to increase stability.
Quality Systems
Our quality systems and processes ensure our success in meeting your needs. ISO 13485 certified facility at Inchinnan, Scotland We are audited by major biopharma and diagnostic companies We employ Six Sigma and Lean Manufacturing to ensure top quality and lower costs Our stringent management controls, process controls, and built-in process improvement steps ensure we meet the quality standards that you expect. We aim to meet quality and regulatory standards and to exceed customer expectations in all that we do. At Invitrogen, we welcome customer and supplier audits as a way to ensure our quality standards meet the expectations and ever-changing needs of our customers.
Product dating
Please refer to product labels for expiration dates of products.
Storage recommendations
For sustained optimal performance, store cell culture products under the conditions recommended on the labels, and minimize exposure of liquid media to all light sources under both storage and usage conditions.25 Keep powder formulations in the dark and dry.
Quality Control
Liquid media
To ensure that our validated systems and controls continue to be effective, the quality of our liquid media is confirmed by testing samples from each production batch. Chemical checks. Osmolality and pH are measured to verify compliance with product specifications. Analytical component testing. Some media products and supplements are tested using analytical techniques like HPLC to verify that contents of the media formulation have been delivered within acceptable limits to assure product performance. Microbiological tests. For liquid media, the absence of bacterial and fungal contamination is confirmed using the method referenced in the current edition of the U.S. Pharmacopeia sterility test monograph. Also, products of animal origin are tested for the absence of mycoplasmas using the large-volume inoculation method of Barile and Kern.1 For details of these tests, see Chapter 4, Microbiological Tests. Growth performance and cytotoxicity tests. Many liquid media are tested for the absence of cytotoxicity and for growth promotion in a suspension culture assay. A rapidly growing murine myeloma cell line, Sp2/0-Ag14, is
Certificates of Analysis
Certificates of Analysis for all cell culture products are available upon request. These documents supplement our standard label data and published specifications with the results of testing performed on all gibco cell culture liquid media and reagents. Also provided is important information about intended product use in keeping with published FDA guidelines.
References
1. 2. 3. 4. Barile, M.F. and Kern, J. (1971) Proc. Soc. Exp. Biol. Med. 138, 432. Wang, R.J. (1976) In Vitro 12, 19. Wang, R.J. and Nixon, B.T. (1978) In Vitro 14, 775. Spierenburg, G.T., Oerlemans, F.T., VanLaarhoven, J.P., and de Bruyn, C.H. (1984) Cancer Res. 44, 2253. 5. Parshad, R., Taylor, W.G., Sanford, K.K., Camalier, R.F., Gantt, R., and Tarone, R.E. (1980) Mutat. Res. 73, 115.
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Introduction
Ordering and Technical Support

For your convenience, you may place your order by the method that is best for you: choose to order online, by email, phone, mail, or fax. Our highly efficient Customer Service representatives will process your order quickly and accurately.
Technical/Customer Support
Our Technical Support lines are open from 6:00 a.m. to 5:00 p.m. Pacific Standard Time. Customer Service representatives are here to assist you with ordering from 5:00 a.m. to 5:00 p.m. Pacific Standard Time. Contact us from anywhere in the US: Technical Support email technical enquiries . . .eurotech@invitrogen.com Email general enquiries . . . . .euroinfo@invitrogen.com Online . . . . . . . . . . . . . . . . . . . . .www.invitrogen.com/support Customer Service email . . . . . . . . . . . . . . . . . . . . . .For specific country email addresses, refer to the back of the catalogue
Order Online
You have two options for placing orders electronically: Place your order online 24 hours a day, 7 days a week, at our website: www.invitrogen.com Email your order: For specific country email addresses, refer to the back of the catalogue
Order by Phone, Mail, or Fax

When you order, please include the following information: 1. Institution or company customer number 2. Billing address 3. Shipping address 4. Valid Purchase Order number 5. Name of purchasing agent and department 6. Name and phone number of end user 7. Catalogue number and description of product 8. Quantity and size of product
Need order status or want to track a shipment? Have a question or need help with an order?
Go to www.invitrogen.com/support and take advantage of our convenient new customer support tools, any time of the day or night.
International Orders
International customers should order directly from the nearest authorized distributor of Invitrogen products. For the location of the nearest distributor, refer to the distributor address list on the inside back cover of this catalogue or at www.invitrogen.com. Where no distributor is designated, orders may be placed with Invitrogen Corporation directly. Shipments outside the continental United States are sent air prepaid. Direct international orders shall require prepayment, payable in U.S. funds via bank wire transfer, Visa, Mastercard, American Express, or the American Express Corporate Purchasing Card. Freight charges will be collected whenever possible.
EUROPEAN HEADQUARTERS
Life Technologies 102 Fountain Crescent Paisley, Scotland PA49RE United Kingdom www.invitrogen.com
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Introduction
Notes:
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Introduction
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .i Resources web links . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i Cell Culture Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii Ordering and Technical Support. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iv
Chapter 1 PSCS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Gibco Cell Therapy Systems (CTS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Gibco CTS Immunotherapy products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 Gibco CTS products for stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 CELLstart Cell Attachment/Expansion Substrate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 Geltrex Reduced Growth Factor Basement Membrane Matrix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Collagen 1 Bovine and Rat Tail Collagen I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 AlgiMatrix 3D Culture System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Natural Mouse Laminin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Chapter 2 Growth factors, Cytokines and Chemokines with high bioactivity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Growth factors, Cytokines and Chemokines with high bioactivity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Gibco Recombinant human FGF-basic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 Gibco LIF Recombinant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 new Gibco Recombinant Growth Factors for use in stem cell applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Human proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 Rat proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 Swine proteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 Publication References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Chapter 3 Basal Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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Reduced-Serum Media at a Glance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 GlutaMAX Media at a Glance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 HEPES Media at a Glance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 Phenol Redfree Media at a Glance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Basal Medium Eagle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 BGJb Medium (Fitton-Jackson Modification) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Brinsters BMOC-3 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Cryopreservation Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 CMRL Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 CO2-Independent Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Dulbeccos Modified Eagle Media (DMEM) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 F-10 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 F-12 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Glasgow Minimum Essential Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Graces Insect Cell Culture Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Improved MEM Zn++ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 IPL-41 Insect Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Iscoves Modified Dulbeccos Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Leibovitzs L-15 Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 McCoys 5A Medium (Modified) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 MCDB 131 Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 Media 199 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 Minimum Essential Media (MEM) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 Advanced MEM (Minimum Essential Medium). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
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Table of Contents
Minimum Essential Medium (MEM) with Earles Salts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 Minimum Essential Medium (MEM) with Hanks Salts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Modified Eagle Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Medium NCTC-109 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 F-10 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 F-12 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Opti-MEM I Reduced-Serum Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 Lipofectamine 2000 Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 RPMI Media 1640 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Schneiders Drosophila Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Waymouths MB 752/1 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Williams Medium E . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Media Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 E-Z Hold Plastic Bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 Stak Pack Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 Media Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Chapter 4 Serum-free and Specialty Media and Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
83
Summary of Serum-Free, Protein-free, and Chemically Defined Media. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Summary of Specialty Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Serum-free Media and Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86 Specialty Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87 3D Matrices and Scaffolds. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88 Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Cellular Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .109 Stem Cell Research Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .117 Cytogenetics Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .126 Overview of Transfection, Selection, and Protein Delivery Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .131 Transfection Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .132
Chapter 5 Balanced Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .137

Balanced Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .138 Dulbeccos Phosphate-Buffered Salines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .138 Earles Balanced Salt Solution. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .139 Hanks Balanced Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .140 Phosphate-Buffered Saline (PBS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .141 Balanced Salt Solutions Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .142 Balanced Salt Solutions Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .143
Chapter 6 Standard Cell Culture Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .147

Cell Culture Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .148 new ACK Lysing Buffer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 Amino Acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 Antibiotics and Antimycotics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .150 Antifoaming and Defoaming Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .156 Biological Buffers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .157 Cell Dissociation Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .158 Distilled Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .161 new WFI for Cell Culture (Water for Injection) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .161 Attachment Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .162 Growth Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .163
viii
Hybridoma Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .166 Insect Medium Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .167 Mycoplasma Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .167 Other Cell Culture Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .168 Cell Culture Reagent Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .169 new Presto Blue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .170
Chapter 7 Eukaryotic Cell Lines
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .173
Insect Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .174 Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .177 Cell Line Development Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .180 new OptiCHO Express Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .180 new OptiCHO Antibody Express Kit. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .181 new Gibco Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .182
Chapter 8 Animal Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .185

Fetal Bovine Sera (FBS) and other Sera for Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .186 FBS Selection Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .187 Fetal Bovine Sera (FBS). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .187 Specialty FBS Selection Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .189 Other Sera Selection Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .190 Newborn Calf Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .191 Bovine Sera. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .191 Horse Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .192 Other Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .192 FBS Quality Control Test Comparison. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .193 Other Sera Quality Control Test Comparison . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .194 Final Product Quality Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .196 Performance Tests: Fetal Bovine Sera (FBS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .197 Special Proccessing of Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .200
Appendix I
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .203 Gibco Universal Bags . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .204 new Lab Armortm Beads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .208 PD-Directtm Bioprocess Services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .209 Cell Culture basics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .210 new Supply Centre by Life Technologiestm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .213 Formulas and Formula Weights for Commonly Used Medium Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .216 Troubleshooting: Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .218 Useful Numbers for Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .219 Buffer Systems for Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .219 Specialty Media and Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .219 Serum-free Media and Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .220 Adaptation of Cell Cultures to a Serum-free Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .224 Adaptation of Cell Cultures to an Alternative Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .225 Adaptation of Cells from Monolayer to Suspension Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .225 Stability of GlutaMAX-I vs. L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .226 Cryopreservation of Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .227
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Thawing of Cryopreserved Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .227 Dissociation of Cells from Primary Tissue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .228 Dissociation of Cells from Culture Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .229 Decontaminating Cultures with Antibiotics and Antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .231 Use of Antibiotics and Antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .231 Conversion Factors, Units, and Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .232 Genetic Code and Amino Acid Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .233
Appendix II Index
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .235 Terms and Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .236 Trademark Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .238 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .239 Alphabetical Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .240 Catalogue Number Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246
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Chapter 1 PSCS
Gibco Cell Therapy Systems (CTS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Gibco CTS Immunotherapy products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 AIM V Medium CTSTherapeutic-grade, serum-free cell expansion medium . . . . . . . . . . . . . 3 OpTmizer CTS T-Cell Expansion SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 CTS cytokines and growth factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 Gibco CTS products for stem cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 CELLstart CTS Cell Attachment/Expansion Substrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 Gibco Extracellular Matrices. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Geltrex Reduced Growth Factor Basement Membrane Matrix . . . . . . . . . . . . . . . . . . . . . . . . 5 Collagen 1 Bovine and Rat Tail Collagen 1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 AlgiMatrix 3D Culture System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 Natural Mouse Laminin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
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Chapter 1 PSCS
Gibco Cell Therapy Systems (CTS)

As you move your translational stem cell, tissue engineering, and immunotherapy research toward the clinic, high-quality products and proper documentation are essential to getting it right the first time. The gibco Cell Therapy Systems (CTS) brand offers a broad array of highquality products designed for use in cell therapy applications, including media, reagents, growth factors, enzymes, selection beads, and devices which are manufactured in compliance with 21 CFR Part 820 Quality System Regulation and/or are certified to ISO 13485 and ISO 9001. CTS products are supplied with unified documentation such as Certificates of Analysis Certificates of Origin Access to authorization letters for our FDA Drug Master File Effective cell therapies require quality raw materials, and that is what you can expect from CTS. The CTS product line will help reduce your burden in qualifying reagents as you prepare to transition your research into clinical applications. The FDA requires manufacturers of cell therapy products to qualify raw materials (reagents, media, growth factors, etc.) used in cell therapy manufacturing processes. Relevant CTS products are mycoplasma-, endotoxin-, and sterility-tested to meet these regulations. In addition, raw materials are USP/EP grade, or an equivalent grade when available. Products labeled RUO or not manufactured under GMPs can create a challenge for the FDA, as performance standards or quality attributes may not have been established. To mitigate these issues, we have established Device Master Files (DMFs) for many of our CTS products to support product development activities. For more information on CTS products, visit www.invitrogen.com/ gibcocts or email us at celltherapysystems@lifetech.com.
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Gibco CTS Immunotherapy products

1
Whether youre developing cell-based vaccines, gene-modified T cells, or other immune-based cell therapy strategies, gibco CTS delivers tools and reagents to support all aspects of your workfrom bench to bedside and beyondinto commercial applications. Gibco CTS provides specialized products to grow, isolate, and expand T cells, dendritic cells, and other cell types. These products are designed to facilitate the development of cell-based vaccines, gene-modified T cells, or other immunity-based cell therapy strategies and to ease your transition into the clinic.
PSCS
AIM V Medium CTSTherapeutic-grade, serum-free cell expansion medium

Gibco AIM V Medium CTS (Therapeutic Grade) is the first commercially available defined, serum-free formulation for proliferation and/or manipulation of T cells and dendritic cells and is manufactured in compliance with cGMP. AIM V Medium CTS is an FDA 510(k)cleared device which is intended for human ex vivo tissue and cell culture processing applications. CAUTION: When used as a medical device, federal law restricts this device to sale by or on the order of a physician.
CTS cytokines and growth factors

With highly active, highly pure gibco CTS cytokines and growth factors, you can culture, expand, and differentiate cell populations with more confidence in your results. Gibco CTS cytokines and growth factors are manufactured under ISO13485 quality standards, and are validated with gibco CTS media, further reducing variability to enable reliable results. For research use or non-commercial manufacturing of cell-based products for clinical research. CAUTION: Not intended for direct administration into humans or animals.
OpTmizer CTS T-Cell Expansion SFM

Gibco OpTmizer CTS T Cell Expansion SFM has been developed for the growth and expansion of human T lymphocytes. OpTmizer CTS T-Cell Expansion Medium is a complete serum-free, xeno-free 1X medium consisting of OpTmizer T Cell Expansion Basal Medium and the OpTmizer T-Cell Expansion Supplement. For research use or manufacturing of cell-, gene-, or tissue-based products. CAUTION: Not intended for direct administration into humans or animals.
Ordering Information
Product AIM VR Medium CTS AIM VR Medium CTS Dynabeads CD3/CD28 CTS DynaMag CTS OpTmizer CTS T-Cell Expansion SFM Interleukin 4 CTS (IL-4) Recombinant Human Interleukin 4 CTS (IL-4) Recombinant Human GM-CSF CTS Recombinant Human GM-CSF CTS Recombinant Human TNF- CTS Recombinant Human TNF- CTS Recombinant Human Interleukin 7 CTS (IL-7) Recombinant Human Interleukin 7 CTS (IL-7) Recombinant Human Interleukin 2 CTS (IL-2) Recombinant Human Interleukin 2 CTS (IL-2) Recombinant Human DPBS CTS (without Calcium Chloride, without Magnesium Chloride) DPBS CTS (with Calcium Chloride, with Magnesium Chloride) 1L A12858-01 Quantity 1,000 mL 10 L 10 mL each 1L 1 mg 100 g 1 mg 100 g 1 mg 100 g 1 mg 100 g 1mg 100 g 1L Cat. no. 087-0112DK 087-0112BK 402-03D 12102 A10485-01 CTP0043 CTP0041 CTP2013 CTP2011 CTP3013 CTP3011 CTP0073 CTP0071 CTP0023 CTP0021 A12856-01
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Chapter 1 PSCS
Gibco CTS products for stem cells

Regardless of the type and source of your stem cells, gibco CTS provides tools to help you isolate, expand, differentiate, and characterize your stem cells. Enter the clinic knowing that your tailored solutions will meet your regulatory and quality requirements.
CELLstart CTS
CELLstart CTS Cell Attachment/ Expansion Substrate
CELLstart substrate contains only components of human origin (xenofree) and supports human embryonic stem cell (hESC) and induced pluripotent stem cell (iPSC) attachment, expansion of undifferentiated colonies, maintenance of pluripotency, and differentiation capability in serum-free medium without the need for feeder cells. CELLstart substrate can also be used with mesenchymal and neural stem cells, and for the attachment of human feeder cells (e.g., foreskin fibroblasts).
Product CELLstart CTS DPBS CTS (without Calcium Chloride, without Magnesium Chloride) DPBS CTS (with Calcium Chloride, with Magnesium Chloride) GlutaMAX-I CTS KnockOut SR XenoFree CTS Kit KnockOut SR XenoFree CTS KnockOut SR XenoFree CTS Kit KnockOut SR XenoFree CTS KnockOut D-MEM CTS TrypLE Select CTS Interleukin 6 CTS Recombinant Human Interleukin 6 CTS Recombinant Human FGF basic full length CTS Recombinant Human FGF basic full length CTS Recombinant Human 1L 100 mL 100 mL 100 mL 500 mL 500 mL 500 mL 100 mL 100ug 1mg 100ug 1mg A12858-01 A12860-01 A1099201 12618-012 A1099202 12618-013 A12861-01 A12859-01 CTP0061 CTP0063 CTP0261 CTP0263 CTP2111 CTP2113 CTP9211 CTP9213 CTP9411 CTP9413 1L A12856-01 Quantity 2 mL Cat. no. A10142-01
Stem Cell Factor (SCF) CTS Recombinant Human 100ug Stem Cell Factor (SCF) CTS Recombinant Human 1mg TGF beta 1 CTS Recombinant Human TGF beta 1 CTS Recombinant Human Flt-3 Ligand CTS Recombinant Human Flt-3 Ligand CTS Recombinant Human 100ug 1mg 100ug 1mg
For Research Use or Non-Commercial Manufacturing of Cell-Based Products for Clinical Research. CAUTION: Not intended for direct administration into human or animals.
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Gibco Extracellular Matrices

1
Geltrex Reduced Growth Factor Basement Membrane Matrix
Geltrex matrix is a soluble form of reduced growth factor (RGF) basement membrane extract (BME) purified from continuous sheets of specialized extracellular matrix that form an interface between Engelbreth-Holm-Swarm (EHS) tumor cells. The major components of Geltrex matrix include laminin, collagen IV, entactin, and heparin sulfate proteoglycan. Geltrex reduced growth factor basement membrane matrix supports cells and cell layers, and plays an essential role in tissue organization that affects cell adhesion, migration, proliferation, and differentiation. Geltrex matrix enables, on average, a 2-fold reduction in several growth factors.
Bovine and Rat Tail Collagen I

Collagen I is a heterotrimeric extracellular matrix protein found in skin and other connective tissues. Gibco Bovine and Rat Tail Collagen I for cell culture are high-quality forms of collagen widely used in 3D applications and as coating solutions. Rat Tail Collagen I is provided in liquid form as well as precoated on plates in 6-well, 24-well, and 96-well formats. Rat Tail Collagen I is the most common fibrillar collagen (90%) found in skin, bone, tendons, and other connective tissues. The liquid form of Rat Tail Collagen I is supplied at 5 mg/mL in a 20 mL vial, providing the flexibility to dilute it to lower concentrations. Bovine Collagen I is a naturally occurring extracellular matrix found in tendons and other connective tissues. It plays a key role in cell growth, differentiation, attachment, and migration. This solution is supplied at 5 mgmL in a 10 mL vial, providing the flexibility to dilute it to lower concentrations.
PSCS
Product Geltrex Reduced Growth Factor Basement Membrane Matrix Geltrex Reduced Growth Factor Basement Membrane Matrix Geltrex LDEV Free Reduced Growth Factor Basement Membrane Extract Geltrex hESC-Qualified Reduced Growth Factor Basement Membrane Matrix Geltrex hESC-Qualified Reduced Growth Factor Basement Membrane Matrix Quantity 1 mL 5 mL 5 mL Cat. no. 12760013 12760021 A1134301
Product Collagen I, Bovine Collagen I, Rat Tail Coated Plates, Collagen I, Rat Tail Coated Plates, Collagen I, Rat Tail Coated Plates, Collagen I, Rat Tail Quantity 10 mL 20 mL 5 x 6-well plates 5 x 24-well plates 5 x 96 well plates Cat. no. A1064401 A1048301 A1142801 A1142802 A1142803
1 mL 5 mL
A1048001 A1048002
For Research Use Only. Not intended for animal or human therapeutic or diagnostic use.
Related products Page HuMEC-SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .114 KnockOut Serum Replacement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .120 Phosphate-Buffered Saline (PBS) 7.2 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . .141 TrypLE Express Stable Trypsin Replacement Enzyme, without phenol red . . . . . . . . . . . . . . . . . . . . . . . . . .160 Versene 1:5,000 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .161 Visit us at www.invitrogen.com/geltrex to learn more about the Geltrex 3D Culture System.
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Chapter 1 PSCS
AlgiMatrix 3D Culture System

The AlgiMatrix 3D Culture System is a user-friendly, nonanimalderived bioscaffold for three-dimensional (3D) cell culture. The AlgiMatrix sponge is formulated using pharmaceutical-grade raw material from brown seaweed. The AlgiMatrix 3D Culture System creates a cell culture model system that more closely reflects normal cell morphology and behavior, to help meet researchers requirements for 3D cell culture in fields such as toxicology, drug development, cancer and stem cell research, development and morphogenesis, tissue and organ engineering, heart disease, and diabetes. The AlgiMatrix system is available in 6-well, 24-well, and 96-well formats that fit well into the workflows of both low- and highthroughput users. Cells can be inoculated directly into the sterile microtiter plates preloaded with lyophilized alginate sponge, with no pretreatment or preparation required. Recommended storage conditions: room temperature.
Natural Mouse Laminin

Natural Mouse Laminin is isolated from Engelbreth-Holm-Swarm (EHS) sarcoma cells and is the major glycoprotein component of basement membranes. Among the functions it modulates are cell attachment, cell spreading, cell growth and motility, promotion of epithelial differentiation, modification of leukocyte function, and stimulation of neurite outgrowth.
Product Natural Mouse Laminin Quantity 1 mg Cat. no. 23017-015
For Research Use Only. Not intended for animal or human therapeutic or diagnostic use.
Product AlgiMatrix 3D Culture System, in 96-well plate AlgiMatrix 3D Culture System, in 96-well plate AlgiMatrix 3D Culture System AlgiMatrix 3D Culture System AlgiMatrix 3D Culture System, 6-well kit AlgiMatrix 3D Culture System, 6-well kit AlgiMatrix Firming Buffer AlgiMatrix Dissolving Buffer Quantity 1 x 96-well plate 5 x 96-well plates 1 x 24-well plate 4 x 24-well plates 1 x 6-well plate 4 x 6-well plates 50 mL 50 mL Cat. no. 12684015 12684031 12684023 12684049 A1098201 A1098202 A1091501 A1134001
For Research Use Only. Not intended for animal or human therapeutic or diagnostic use. Related products Page Defined Keratinocyte-SFM System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .110 Hepatozyme-SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .113 KnockOut Serum Replacement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .120 Phosphate-Buffered Saline (PBS) 7.2 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . .141 TrypLE Express Stable Trypsin Versene 1:5,000 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .160 Visit www.invitrogen.com/3D-CellCulture to learn more about the AlgiMatrix 3D Culture System and related products for 3D cell culture.
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Chapter 1 PSCS
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Growth factors, Cytokines and Chapter 2 Chemokines with high bioactivity

Growth factors, Cytokines and Chemokines with high bioactivity. . . . . . . . . . . . . . . . . . . . . . . . . 10 Gibco Recombinant human FGF-basic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 Gibco LIF Recombinant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 new Gibco Recombinant Growth Factors for use in stem cell applications . . . . . . . . . . . . . . . . . 14 Thrombopoietin (TPO) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 BMP-4 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 BMP-7 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Activin A . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 IL-17F . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 Human proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 Rat proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 Swine proteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 Publication References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
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Chapter 2 Growth factors, Cytokines, and Chemokines with high bioactivity
Growth factors, Cytokines and Chemokines with high bioactivity

Proven Compatibility
gibco recombinant proteins have proven compatibility with gibco media. gibco recombinant proteins are bioassayed with gibco media. You can be confident that your recombinant protein is compatible with the media you trust.
High Quality
All gibco recombinant proteins from our Camarillo, CA, and Frederick, MD, sites are manufactured under ISO 13485 quality standards. These standards require that all proteins deliver high bioactivity, low endotoxin, and consistent performance.
High Biological Activity

gibco recombinant proteins have biological activity that meets or exceeds industry standards. Higher activity provides more results with less protein.
Multiple Packaging Sizes

Many gibco recombinant proteins are available in multiple package sizes to meet your application needs. Buffer formulations and package sizes may be customized upon request. For more information, please contact Custom Services at evoquestteam@invitrogen.com or by phone at 800 955 6288.
High Purity
gibco recombinant proteins are highly purified. High purity means no interference from other proteins or contaminant. When working with gibco recombinant proteins you get results you can trust.
How are your cytokines, chemokines, and growth factors manufactured?

Process
Expression Gene cloned from cDNA Sequence verified Expression system chosen Purification and refolding Purification by chromatography Refolding of proteins expressed in bacterial system Formulation and lyophilization
Carrier-free Sterile-filtered prior to

lyophization
Result
Expressed in a host system that yields a biologically active protein
Purified to produce a highly pure, homogenous solution of active protein
formulated and lyophilized to ensure maximum stability
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How is the quality of your growth factors ensured?

To ensure gibco growth factors are of the highest quality, each protein is analyzed for purity along with structural homogeneity to ensure a biologically active protein is produced. In-house bioactivity testing includes cell proliferation, cytotaxis, calcium flux, secondary cytokine up-regulation, induction of surface antigen expression, and protease assays.
Characteristic Purity Refolding Structural homogeneity Biological activity Stability Endotoxin Test performed SDS-PAGE Bioassay SDS-PAGE Published bioassay Freeze/thaw LAL (Limulus amoebocyte lysate) method Criteria for success >95% purity Bioactive protein Clean band ED50 other available vendors Negligible loss <0.1 g/mg Figure 1 2 3 4, 5, 6
2 Growth factors, Cytokines and Chemokines with high bioactivity
RANTES
SCF
1.2 1.1 1.0
ED50 (pg/mL) GIBCO = 548.4 Competitor = 6593.6
Mean OD
0.9 0.8 0.7 0.6 0.5 0.4 0 101 102 103 104 105 106
[SCF] (pg/mL)
Figure 1. High purity. SDS-PAGE analysis of purified recombinant human RANTES (Cat. No. PHC1054) and mouse SCF (Cat. No. PMC2114).
Figure 2. More bioactive protein due to exceptional refolding techniques. Proliferation of MO7e cells in response to SCF from gibco and the competitor (Cat. No. PHC2115). As illustrated by the lower ED50, less gibco SCF is required to yield a response.
Figure 3. Structural homogeneity. The SDS-PAGE gel was run on Human BMP-2 (Cat. No. PHC7145) under reducing (lanes 2 and 3) and non-reducing (lanes 4 and 5) conditions. Under non-reducing conditions, BMP-2 is seen as a dimer, consistent with biologically active BMP-2.
2.4
2.2 2.0 1.8
Fluorescent intensity
2.2
ED50 (ng/mL) GIBCO = 0.26 Competitor = 0.58
280,000 240,000 200,000
RFU
2.0
Mean OD
1.6 1.4 1.2
160,000 120,000 80,000 40,000 0 0

Substrate only Competitors human KLK4 GIBCO Human KLK4
1.8
1.6
1.0 0.8 0.001 0.01 0.1 1.0 10 100
1.4 290 300
320
340
360
380
400
1,000 2,000 3,000 4,000 5,000 6,000 7,000
Time (sec)
[FGF] (ng/mL)
Time (sec)
Figure 4. Calcium flux confirms expected biological response. Calcium flux activation was observed by stimulating Fura-2loaded rat PMNs with the gibco chemokine CINC-2 (Cat. No. PRC1564).
Figure 5. High bioactivity of FGF-basic. Proliferation of Balb/3T3 cells in response to FGF (Cat. No. PHG0024) is demonstrated. gibco FGF-basic is more active than the competitors FGF-basic.
Figure 6. Protease activity confirms protein functionality. Enzymatic activity of Kallikrein 4 (KLK4) was measured by its ability to cleave fluorogenic peptide substrate Boc-Val-ProArg-AMC. The end product is measured using fluorescence excitation and emission wavelengths of 380 nm and 460 nm, respectively.
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Gibco Recombinant human FGF-basic

FGF-basic
FGF-basic is a critical component for maintaining embryonic stem cells in culture in an undifferentiated state. gibco offers recombinant human FGF-basic, a bioactive protein intended for use in cell culture applications. gibco comes in two forms: 1) FGF-basic (AA 1-155)a full-length version 2) FGF-basic (AA 10-155)a truncated version Youve trusted gibco to provide high quality cell culture media and reagents. Now supplement your media with gibco FGF-basic!
Figure 1Balb/3T3 cells (mouse fibroblast cell line, ATCC: CCL-163) at constant density were co-cultured with serial dilutions of FGF-basic samples in serumfree medium at 37 oC, 5% CO2 between 18 24 hours. The optical density (OD) of each concentration of the testing samples, which is directly proportional to the cell number, was read at 490/650 nm at the end point of incubation. The cell proliferation dose curve was plotted with mean OD vs. FGF-b concentrations (ng/ml).
Figure 2ED50 of Recombinant human FGF-basic (AA 10-155). The optical density (OD) of each concentration of the testing samples, which is directly proportional to the cell number, was read at 490/650 nm at the end point of incubation. The cell proliferation dose curve was plotted with mean OD vs. FGF-b concentrations (ng/ml).
Product Name FGF-basic (AA 1-155) Recombinant Human FGF-basic (AA 1-155) Recombinant Human FGF-basic (AA 1-155) Recombinant Human FGF-basic (AA 1-155) Recombinant Human FGF-basic (AA 10-155) Recombinant Human FGF-basic (AA 10-155) Recombinant Human FGF-basic (AA 10-155) Recombinant Human FGF-basic (AA 10-155) Recombinant Human FGF-basic (AA 10-155) Recombinant Human (Liquid Form) Species Human Human Human Human Human Human Human Human Human Quantity 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 50 g 100 g Cat. no. PHG0261 PHG0263 PHG0264 PHG0266 PHG0021 PHG0023 PHG0024 PHG0026 PHG0021L
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Gibco LIF Recombinant

LIF Recombinant
While known for its pleiotropic actions, Leukemia Inhibitory Factor (LIF) is most notable for its ability to prevent the differentiation of mouse embryonic stem cells in culture enabling them to maintain their pluripotency. gibco offers both recombinant mouse LIF and recombinant human LIF, bioactive proteins intended for use in cell culture applications. Youve trusted gibco to provide high quality cell culture media and reagents. Now supplement your media with gibco LIF! Mouse embryonic stem cells cultured with gibco recombinant LIF maintain pluripotency Pluripotency of mouse embryonic stem cells cultured with gibco LIF has been confirmed by alkaline phosphatase expression, by expression of pluripotency markers, and by the ability of the mouse embryonic stem cells to generate the three germ layers. Alkaline phosphatase is a marker of undifferentiated embryonic stem cells Biological activity of gibco LIF
2.9 2.7 2.5
Experiment#1 Recombinant LIF
2.3 2.1 1.9 1.7 1e-4
Growth factors, Cytokines and Chemokines with high bioactivity
0.001
0.01
0.1 Concentration (ng/ml)
10
100
4-P B C D R^2 Figure 2The Fit: y = (A - D)/( 1 + (x/C)^B ) + D: A of MTS within stimulated mouse M1 dose dependent reduction Plot#1 (Competitor: Concentration vs MeanValue) 1.78 1.81 0.253 2.8 0.989 cells to measure the biological activity of a) gibco recombinant mouse LIF b) Plot#2 (GIBCO: Concentration vs MeanValue) 1.78 1.39 0.138 2.82 0.999 __________ gibco recombinant human LIF compared to the competitor LIF. Weighting: Fixed
Product LIF Recombinant Human LIF Recombinant Human LIF Recombinant Mouse LIF Recombinant Mouse Quantity 100 g 10 g 100 g 10 g Cat. no. PHC9461 PHC9464 PMC4051 PMC4054
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Figure 1a) undifferentiated mouse embryonic stem cells and; b) differentiated mouse embryonic stem cells; and c) mouse embryonic stem cells cultured in 15% KnockOut TM Serum Replacement (KSR) and 10 ng/mL of gibco mouse LIF.
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new
Gibco Recombinant Growth Factors for use in stem cell applications
We are expanding our line of gibco recombinant proteins with three growth factors important to stem cell researchThrombopoietin (TPO), BMP-4, and Activin-A. These join other gibco growth factors and cytokines used in stem cell research, like GM-CSF, TGF, FGF basic, Stem Cell Factor, TGF-1, Flt-3 Ligand, IL-2, IL-4, IL-6, and IL-7.
Thrombopoietin (TPO)
TPO is a humoral growth factor that is necessary for megakaryocyte proliferation and maturation, as well as for thrombopoiesis. This protein is the ligand for MLP/MPL, the product of myeloproliferative leukemia virus oncogene.
Product Name Thrombopoietin, Recombinant Human Thrombopoietin, Recombinant Human Thrombopoietin, Recombinant Human Species Human Human Human Quantity 10g 100g 1 mg Cat. no. PHC9514 PHC9511 PHC9513
BMP-4
BMP4 is a member of the bone morphogenetic protein family which is part of the transforming growth factor-beta superfamily. Bone morphogenetic proteins were originally identified by an ability of demineralized bone extract to induce endochondral osteogenesis in vivo in an extraskeletal site. This particular family member plays an important role in the onset of endochondral bone formation in humans.
Product Name BMP-4, Recombinant Human BMP-4, Recombinant Human BMP-4, Recombinant Human Species Human Human Human Quantity 10 g 100 g 1 mg Cat. no. PHC9534 PHC9531 PHC9533
BMP-7
BMP-7 plays a role in the differentiation of mesenchymal cells into bone and cartilage. BMP7 may also be involved in bone homeostasis.
Product Name BMP-7, Recombinant Human BMP-7, Recombinant Human BMP-7, Recombinant Human Quantity 10 g 100 g 1000 g Cat. no. PHC9544 PHC9541 PHC9543
Activin A
Activin A is a member of the transforming growth factor-beta superfamily. Activin A plays a role in hormone secretion, neuronal survival, and proliferation.
Product Name Activin A, Recombinant Human Activin A, Recombinant Human Activin A, Recombinant Human Species Human Human Human Quantity 10 g 100 g 1 mg Cat. no. PHC9564 PHC9561 PHC9563
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IL-17F
IL-17F is unregulated during the inflammation response. IL-17F has been implicated in the allergic response.
Product Name IL-17F, Recombinant Human IL-17F, Recombinant Human IL-17F, Recombinant Human Quantity 10 g 100 g 1000 g Cat. no. PHC9554 PHC9551 PHC9553
Human proteins
Product 4-1BBL 6CKine/SLC 6CKine/SLC 6CKine/SLC Active Caspase-3 Activin A Activin R1b new Activin R1b new Activin R1b new Activin R1b new -Defensin (NP-1) -Synuclein new Angiopoietin-1 Angiopoietin-2 BAFF BCA-1 (CXCL13) new -Defensin (BD-1) B-DNF -NGF new -Synuclein new BMP-2 new BMP-2 new BMP-2 new BMP-2 new BMP-4 BMP-7 (active) BMP-7 (inactive) BMPR1A new BMPR1A new BMPR1A new Cardiotrophin-1 Quantity 20 g 10 g 100 g 1 mg 100 units 5 g 10 g 25 g 100 g 1 mg 20 g 200 g 10 g 10 g 20 g 20 g 20 g 10 g 20 g 200 g 10 g 25 g 100 g 1 mg 5 g 10 g 10 g 10 g 100 g 1 mg 10 g Cat. no. PHC1564 PHC1474 PHC1471 PHC1473 PHZ0014 PHG9014 PHC9284 PHC9285 PHC9281 PHC9283 PHC1615 PHB0044 PHC1514 PHC1524 PHC1674 PHC0052 PHC1624 PHC7074 PHG0126 PHB0064 PHC7145 PHC7146 PHC7141 PHC7143 PHC7914 PHC7204 PHC7104 PHC9244 PHC9241 PHC9243 PHC1594 Product sCD4 new CD40 Ligand/TRAP (soluble) CD40 Ligand/TRAP (soluble) CNTF Cripto new Cripto new Cripto new Cripto new CTGF CTLA-Fc Dkk-1 new Dkk-1 new Dkk-1 new Dkk-1 new EGF new EGF new EGF new EGF new EGF (liquid) new EMAP-II ENA-78 ENA-78 ENA-78 ENA-78 Eotaxin Eotaxin Eotaxin Erythropoietin Quantity 10 g 10 g 50 g 20 g 10 g 25 g 100 g 1 mg 20 g 100 g 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg 100 g 20 g 10 g 25 g 100 g 1 mg 10 g 100 g 1 mg 500 IU Cat. no. PHS0044 PHP0024 PHP0025 PHC7015 PHC8014 PHC8015 PHC8011 PHC8013 PHG0286 PHR5001 PHC9214 PHC9215 PHC9211 PHC9213 PHG0314 PHG0315 PHG0311 PHG0313 PHG0311L PHC1585 PHC1335 PHC1336 PHC1331 PHC1333 PHC1434 PHC1431 PHC1433 PHC2054 Product FGF-4 FGF-6 FGF-7 FGF-8 FGF-8b FGF-8b FGF-8b FGF-8b FGF-10 FGF-16 FGF-17 FGF-18 FGF-19 FGF-9 FGF-acidic FGF-basic FGF-basic FGF-basic FGF-basic aa10155 (liquid) new FGF-basic FGF-basic (full length) FGF-basic (full length) FGF-basic (full length) FGF-basic (full length) Flt-3 Ligand new Flt-3 Ligand new Flt-3 Ligand new Flt-3 Ligand new Quantity 25 g 25 g 10 g 25 g 10 g 25 g 100 g 1 mg 25 g 25 g 25 g 25 g 25 g 20 g 10 g 10 g 50 g 100 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg Cat. no. PHG0154 PHG0174 PHG0094 PHG0184 PHG0274 PHG0276 PHG0271 PHG0273 PHG0204 PHG0214 PHG0224 PHG0234 PHG0244 PHG0194 PHG0014 PHG0024 PHG0026 PHG0021 PHG0021L PHG0023 PHG0264 PHG0266 PHG0261 PHG0263 PHC9414 PHC9415 PHC9411 PHC9413
Growth factors, Cytokines and Chemokines with high bioactivity 15
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Human proteins, continued
Product Frizzled-1 new Frizzled-1 new Frizzled-1 new Frizzled-1 new Frizzled-4 new Frizzled-4 new Frizzled-7 new Frizzled-7 new G-CSF G-CSF G-CSF new G-DNF G-DNF G-DNF G-DNF new GM-CSF GM-CSF GM-CSF GRO- GRO- GRO- GRO- GRO- GRO- (CXCL2, AA-39-107) new GRO- (CXCL2, AA-39-107) new HGF IFN- A IFN- A IFN- A/D, Bgl II Hybrid IFN- B2 IFN- D Subtype IFN- new IFN- IFN- IGF-II IL-1 IL-1 IL-1 IL-1 IL-1 IL-1 IL-1 IL-1 IL-1
Quantity 10 g 25 g 100 g 1 mg 10 g 100 g 10 g 100 g 25 g 100 g 1 mg 5 g 10 g 100 g 1 mg 10 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 10 g 100 g 10 g 5 g 105 units 105 units 106 units 10 units 100 g 1 mg 50 g 5 g 25 g 100 g 1 mg 2 g 10 g 25 g 100 g 1 mg
Cat. no. PHC9384 PHC9385 PHC9381 PHC9383 PHC9434 PHC9431 PHC9474 PHC9471 PHC2035 PHC2031 PHC2033 PHC7044 PHC7045 PHC7041 PHC7043 PHC2015 PHC2011 PHC2013 PHC1065 PHC1066 PHC1061 PHC1063 PHC1074 PHC9224 PHC9221 PHG0254 PHC4014 PHC4044 PHC4064 PHC4054 PHC4244 PHC4031 PHC4033 PHG9074 PHC0015 PHC0017 PHC0011 PHC0013 PHC0814 PHC0815 PHC0816 PHC0811 PHC0813
Product IL-2 IL-2 IL-2 IL-2 IL-3 IL-3 IL-3 IL-3 IL-4 IL-4 IL-4 IL-4 IL-5 IL-6 IL-6 IL-6 IL-6 IL-6 IL-7 IL-7 IL-7 IL-7 IL-8 (endothelial) IL-8 (monocyte) IL-8 (monocyte) IL-8 (monocyte) IL-8 (monocyte) IL-10 IL-10 IL-10 IL-10 IL-11 IL-12 new IL-12 new IL-13 IL-13 IL-13 IL-15 IL-15 IL-17 IL-17 IL-17 IL-17 IL-18/IGIF IL-19 IL-19 IL-20 IL-20 IL-20
Quantity 10 g 40 g 100 g 1 mg 10 g 50 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 5 g 10 g 25 g 100 g 1 mg 5 g 25 g 100 g 1 mg 25 g 25 g 50 g 100 g 1 mg 5 g 10 g 100 g 1 mg 10 g 10 g 1 mg 5 g 25 g 100 g 10 g 100 g 10 g 25 g 100 g 1 mg 25 g 10 g 100 g 10 g 25 g 100 g
Cat. no. PHC0026 PHC0027 PHC0021 PHC0023 PHC0034 PHC0035 PHC0031 PHC0033 PHC0044 PHC0045 PHC0041 PHC0043 PHC0055 PHC0064 PHC0065 PHC0066 PHC0061 PHC0063 PHC0075 PHC0076 PHC0071 PHC0073 PHC0084 PHC0884 PHC0885 PHC0881 PHC0883 PHC0104 PHC0105 PHC0101 PHC0103 PHC0115 PHC1124 PHC1123 PHC0134 PHC0135 PHC0131 PHC9154 PHC9151 PHC9174 PHC0174 PHC9171 PHC9173 PHC0186 PHC0194 PHC0191 PHC0204 PHC0205 PHC0201
Product IL-21 IL-21 IL-21 IL-21 IL-29 new IL-29 new IL-31 new IL-31 new IL-33 new IL-33 new IL-33 new IL-33 new IP-10 IP-10 IP-10 IP-10 ITAC ITAC Kallikrein 4 new Kallikrein 4 new Kallikrein 4 new Kallikrein 4 new Kallikrein 7 new Kallikrein 7 new Kallikrein 7 new Kallikrein 7 new Kallikrein 11 new Kallikrein 11 new Kallikrein 11 new Kallikrein 11 new KGF-2 Leptin LIF new LIF new MCP-1 MCP-1 MCP-1 MCP-2 MCP-2 new MCP-2 MCP-2 MCP-3 MCP-4 M-CSF M-CSF MDC/STCP-1 Midkine
Quantity 10 g 25 g 100 g 1 mg 10 g 100 g 10 g 100 g 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 100 g 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg 25 g 1 mg 10 g 100 g 10 g 25 g 100 g 5 g 10 g 25 g 100 g 10 g 10 g 10 g 100 g 20 g 20 g
Cat. no. PHC0214 PHC0215 PHC0211 PHC0213 PHC9274 PHC9271 PHC9264 PHC9261 PHC9254 PHC9255 PHC9251 PHC9253 PHC1084 PHC1085 PHC1081 PHC1083 PHC1694 PHC1691 PHC9344 PHC9345 PHC9341 PHC9343 PHC9294 PHC9295 PHC9291 PHC9293 PHC9314 PHC9315 PHC9311 PHC9313 PHC1536 PHP0013 PHC9464 PHC9461 PHC1014 PHC1015 PHC1011 PHC1114 PHC1116 PHC1115 PHC1111 PHC1574 PHC1154 PHC9504 PHC9501 PHC1204 PHC7084
5 x 106 units PHC4814
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Product MIG MIG MIG MIP-1 MIP-1 MIP-1 new MIP-1 (liquid) MIP-1 (liquid) MIP-3 MIP-3 MIP-4 MIP-5 Natural Fibronectin new Natural Vitronectin new Neurotactin/Fractalkine Neurotrophic Factor-4 (NT-4) Neurturin Neutrophil Activation Protein-2 (NAP-2) Neutrophil Activation Protein-2 (NAP-2) Noggin new Oncostatin M Osteoprotegerin (OPG) PDGF-AA PDGF-AB
Quantity 10 g 1 mg 100 g 10 g 100 g 10 g 1 mg 100 g 20 g 20 g 10 g 25 g 1 mg 100 g 20 g 5 g 10 g 2 g 10 g 20 g 10 g 50 g 10 g 10 g
Cat. no. PHC1604 PHC1603 PHC1601 PHC1104 PHC1101 PHC1034Z PHC1033L PHC1031L PHC1234 PHC1244 PHC1254 PHC1544 PHE0023 PHE0011 PHC1174 PHC7024 PHC7064 PHC1094 PHC1095 PHC1506 PHC5015 PHC1684 PHG0035 PHG0134
Product PDGF-BB PDGF-BB PDGF-BB PDGF-BB PDGF-BB PF4 new PIGF-1 Pleiotrophin (PTN, HARP) RANK Ligand RANTES RANTES RANTES RANTES SCF/c-Kit Ligand SCF/c-Kit Ligand SCF/c-Kit Ligand SCF/c-Kit Ligand SDF-1 new SDF-1 new SDF-1 new SDF-1 new SDF-1 TARC TECK TGF-
Quantity 5 g 10 g 50 g 100 g 1 mg 20 g 20 g 20 g 10 g 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 25 g 100 g 1 mg 10 g 20 g 20 g 100 g
Cat. no. PHG0044 PHG0045 PHG0046 PHG0041 PHG0043 PHC7056 PHG0296 PHC7075 PHP0034 PHC1054 PHC1055 PHC1051 PHC1053 PHC2115 PHC2116 PHC2111 PHC2113 PHC1344 PHC1346 PHC1341 PHC1343 PHC1364 PHC1264 PHC1625 PHG0051
Product TGF-1 TGF-1 TGF-1 new TGF-1 new TGF-1 TGF-2 TGF-3 TGF-3 TIMP-1 new TIMP-1 new TIMP-1 new TNF- TNF- (liquid) TNF- TNF- TNF- TNF-RI TNF-RII TRAIL VEGF new VEGF new VEGF new Wnt-1 new
Quantity 1 mg 5 g 10 g 100 g 250 g 5 g 5 g 250 g 10 g 100 g 1 mg 10 g 10 g 50 g 100 g 1 mg 20 g 20 g 50 g 10 g 100 g 1 mg 10 g
Cat. no. PHG9203 PHG9204 PHG9214 PHG9211
PHG9202 PHG9114 PHG9305 PHG9302 PHC8024 PHC8021 PHC8023 PHC3015 PHC3015L PHC3016 PHC3011 PHC3013 PHR3015 PHR3025 PHC1634 PHC9394 PHC9391 PHC9393 PHC1804
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Rat proteins
Product Ciliary Neurotrophic Factor (CNTF) GM-CSF GM-CSF GM-CSF GRO/KC/CINC-1 GRO-/MIP-2 GRO-/MIP-2 IFN- IFN- IFN- IFN- IFN- IL-1 IL-1 IL-1 IL-1 IL-1 IL-1 IL-2 IL-2 IL-3 IL-3 Quantity 25 g 10 g 25 g 100 g 25 g 100 g 10 g 2 x 105 units 3 x 105 units 10 g 100 g 1 mg 10 g 100 g 10 g 25 g 100 g 1 mg 10 g 100 g 10 g 100 g Cat. no. PRC7015 PRC2015 PRC2016 PRC2011 PRC1066 PRC1071 PRC1074 PRC4014 PRC4024 PRC4034 PRC4035 PRC4033 PRC0014 PRC0011 PRC0814 PRC0815 PRC0811 PRC0813 PRC0024 PRC0021 PRC0034 PRC0031 Product IL-5 IL-5 IL-5 IL-6 IL-6 IL-6 IL-10 IL-13 IL-13 IL-13 IL-18/IGIF IL-18/IGIF MCP-1 MCP-1 MCP-1 RANTES TNF- TNF- TNF- VEGF VEGF VEGF VEGF Quantity 10 g 100 g 1 mg 10 g 100 g 1 mg 10 g 10 g 100 g 1 mg 10 g 100 g 10 g 25 g 100 g 20 g 10 g 100 g 1 mg 10 g 25 g 100 g 1 mg Cat. no. PRC0054 PRC0051 PRC0053 PRC0064 PRC0061 PRC0063 PRC0104 PRC0134 PRC0131 PRC0133 PRC0184 PRC0181 PRC1014 PRC1015 PRC1011 PRC1054 PRC3014 PRC3015 PRC3013 PRG0114 PRG0115 PRG0111 PRG0113
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Swine proteins
Product GM-CSF GM-CSF GM-CSF IFN- IFN- IL-1 IL-1 IL-2 IL-2 IL-4 IL-4 IL-4 IL-6 IL-6 IL-8 IL-8 IL-10 IL-10 IL-15 IL-15 TNF- TNF- Quantity 10 g 25 g 100 g 10 g 100 g 10 g 100 g 10 g 100 g 10 g 25 g 100 g 10 g 100 g 10 g 100 g 10 g 100 g 10 g 100 g 10 g 100 g Cat. no. PSC2014 PSC2015 PSC2011 PSC4034
PSC4031 PSC0014 PSC0011 PSC0024 PSC0021 PSC0044 PSC0045 PSC0041 PSC0064 PSC0061 PSC0884 PSC0881 PSC0104 PSC0101 PSC0154 PSC0151 PSC3014 PSC3011

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Have your cytokines, chemokines, and growth factors been referenced in publications?
Proven Compatibility
Ng F et al. (2008) PDGF, TGF-, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): transcriptional profiling can identify markers and signaling pathways important in differentiation of MSCs into adipogenic, chondrogenic, and osteogenic lineages. Blood 112:295307.
Human IL-21
de Totero B et al. (2008) The opposite effects of IL-15 and IL-21 on CLL B cells correlate with differential activation of the JAK/STAT and ERK1/2 pathways. Blood 111:517524.
Human MIP-1
Bluman EM et al. (1996) Human natural killer cells produce abundant macrophage inflammatory protein-1 alpha in response to monocytederived cytokines. J Clin Invest 97:27222727.
Human EGF
Shimada T et al. (2009) Development of targeted therapy with paclitaxel incorporated into EGF-conjugated nanoparticles. Anticancer Res 29:10091014.
Human PDGF-BB
Hou S et al. (2002) In adenovirus type 12 tumorigenic cells, major histocompatibility complex class I transcription shutoff is overcome by induction of NF-B and relief of COUP-TFII repression. J Virol 76:32123220.
Human FGF-basic
Levenstein M et al. (2006) Basic fibroblast growth factor support of human embryonic stem cell self-renewal. Stem Cells 24:568574. Jerebtsova M et al. (2007) A novel role of fibroblast growth factor-2 and pentosan polysulfate in the pathogenesis of intestinal bleeding in mice. Am J Physiol Heart Circ Physiol 292:743750. Lu S et al. (2008) Biologic properties and enucleation of red blood cells from human embryonic stem cells. Blood 112:44754484. Karumbayaram S et al. (2009) Human embryonic stem cell-derived motor neurons expressing SOD1 mutants exhibit typical signs of motor neuron degeneration linked to ALS. Dis Model Mech 2:189195. Zhou J et al. (2009) mTOR supports long-term self-renewal and suppresses mesoderm and endoderm activities of human embryonic stem cells. Proc Natl Acad Sci U S A 106:78407845.
Human SCF/c-kit ligand

Denis MM et al. (2005) Escaping the nuclear confines: signal-dependent pre-mRNA splicing in anucleate platelets. Cell 122:379391. Lindberg SR et al. (2005) The leukemia-associated ETO homologues are differently expressed during hematopoietic differentiation. Exp Hematol 33:189198.
Mouse GM-CSF
Jamur MC et al. (2005) Identification and characterization of undifferentiated mast cells in mouse bone marrow. Blood 105:42824289.
Human G-CSF
Matsui W et al. (2005) Requirement for myeloid growth factors in the differentiation of acute promyelocytic leukaemia. Br J Haematol 128:853862.
Mouse IL-6
Selander C et al. (2009) TLR2/MyD88-dependent and -independent activation of mast cell IgE responses by the skin commensal yeast Malassezia sympodialis. J Immunol 182:42084216.
Human IL-2
Zeddou M et al. (2005) Prostaglandin E2 induces the expression of functional inhibitory CD94/NKG2A receptors in human CD8+ T lymphocytes by a cAMP-dependent protein kinase A type I pathway. Biochem Pharmacol 70:714724. Kemper C et al. (2003) Activation of human CD4+ cells with CD3 and CD46 induces a T regulatory cell 1 phenotype. Nature 421:388392.
Mouse IL-7
Alter-Wolf S et al. (2009) Deviation of the B cell pathway in senescent mice is associated with reduced surrogate light chain expression and altered immature B cell generation, phenotype, and light chain expression. J Immunol 182:138147.
Mouse PDGF-BB
Clair T et al. (2003) Autotaxin hydrolyzes sphingosylphosphorylcholine to produce the regulator of migration, sphingosine-1-phosphate. Cancer Res 63:54465453.
Human IL-4
Pietrella D et al. (2008) A Candida albicans mannoprotein deprived of its mannan moiety is efficiently taken up and processed by human dendritic cells and induces T cell activation without stimulating proinflammatory cytokine production. Infect Immun 76:43594367.
Mouse SCF
Alter-Wolf S et al. (2009) Deviation of the B cell pathway in senescent mice is associated with reduced surrogate light chain expression and altered immature B cell generation, phenotype, and light chain expression. J Immunol 182:138147.
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Notes:
2 Growth factors, Cytokines and Chemokines with high bioactivity 21

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Notes:
22
Chapter 3 Basal Media

Reduced-Serum Media at a Glance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 GlutaMAX Media at a Glance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 HEPES Media at a Glance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 Phenol Redfree Media at a Glance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Basal Medium Eagle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 BGJb Medium (Fitton-Jackson Modification) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 Brinsters BMOC-3 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Cryopreservation Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 CMRL Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 CO2-Independent Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Dulbeccos Modified Eagle Media (DMEM) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 Advanced DMEM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 DMEM (high glucose) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 DMEM (low glucose). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 DMEM (no glucose) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 DMEM, powder (low glucose) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 DMEM Nutrient Mixture F-12 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 F-10 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 F-12 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Glasgow Minimum Essential Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Graces Insect Cell Culture Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Improved MEM Zn++ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 IPL-41 Insect Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Iscoves Modified Dulbeccos Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Leibovitzs L-15 Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 McCoys 5A Medium (Modified) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 MCDB 131 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 Media 199 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 Minimum Essential Media (MEM) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 Advanced MEM (Minimum Essential Medium) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 Minimum Essential Medium (MEM) with Earles Salts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 Minimum Essential Medium (MEM) with Hanks Salts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
continued
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Modified Eagle Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Medium NCTC-109 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 F-10 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 F-12 Nutrient Mixtures. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 Opti-MEM I Reduced-Serum Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 Lipofectamine 2000 Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 RPMI Media 1640 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Schneiders Drosophila Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Waymouths MB 752/1 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Williams Medium E . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Media Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 E-Z Hold Plastic Bottles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 Stak Pack Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 Media Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
24
Reduced-serum Media at a Glance

Reduced-serum media significantly reduce serum requirements, which helps decrease variability and extend the life of a serum lot. Advanced media formulations reduce FBS supplementation by 5090%, and Opti-MEM Media reduce FBS supplementation by at least 50%, with no change in cell growth, promotion, morphology, or function. For your convenience, this page consolidates all gibco reduced-serum media in a concise, easy-to-use table. You can quickly scan it to determine what you need and where the product is fully described in this catalogue. For performance testing data of Advanced media go to www.invitrogen.com/advanced.
Reduced-serum Media
Product Advanced DMEM (Dulbeccos Modified Eagle Medium) (1X), liquid Advanced DMEM/F-12 (1X), liquid, 1:1 Advanced MEM (Minimum Essential Medium) (1X), liquid Advanced RPMI Medium 1640 (1X), liquid Opti-MEM I Reduced-Serum Medium (1X), liquid Opti-MEM I Reduced-Serum Medium (1X), liquid (w/o phenol red) Opti-MEM I Reduced-Serum Medium (1X), liquid (with GlutaMAX-I) Contains 4,500 mg/L D-glucose, non-essential amino acids (NEAA), and 110 mg/L of sodium pyruvate, but no L-glutamine Contains 3,151 mg/L D-glucose, non-essential amino acids (NEAA), and 110 mg/L sodium pyruvate, but no L-glutamine Contains 110 mg/L of sodium pyruvate and non-essential amino acids (NEAA), but no L-glutamine Contains 2,000 mg/L D-glucose, non-essential amino acids (NEAA), and 110 mg/L sodium pyruvate, but no L-glutamine Contains HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1.1 mg/L Contains HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, and growth factors, but no phenol red Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1.1 mg/L A modification of MEM (Eagles) Contains HEPES buffer, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1.1 mg/L, but no sodium bicarbonate Quantity 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 100 ml 10 x 500 ml (case) Cat. no. 12491-015 12491-023 12634-010 12634-028 12492-013 12492-021 12633-012 12633-020 31985-047 31985-054 Page 68 71
3 Basal Media
78 83 81
500 ml
11058-021
81
500 ml 10 500 ml (case)
51985-026 51985-042
81
Opti-MEM I Reduced-Serum Medium, powder
10 1 L
22600-134
81
For more information on classifications, production, or preparation of the products in this chapter, please see pages 47 and 48.
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GlutaMAX Media at a Glance

GlutaMAX Media have increased stability, minimize toxic ammonia build-up, and improve cell viability. They are standard cell culture formulations that contain L-glutamine as a stabilized dipeptide. For your convenience, this page consolidates all gibco GlutaMAX Media in a concise, easy-to-use table. You can quickly scan it to determine what you need and where the product is fully described in this catalogue.
GlutaMAX Media
Product Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid DMEM/F-12, (1X) liquid, 1:1 F-12 Nutrient Mixture (Ham) (1X), liquid Iscoves Modified Dulbeccos Medium (IMDM) (1X), liquid Minimum Essential Medium (MEM) alpha (1X), liquid Minimum Essential Medium (MEM) alpha (1X), liquid Minimum Essential Medium (MEM), liquid Minimum Essential Medium (MEM), liquid Opti-MEM I Reduced-Serum Medium (1X), liquid RPMI Medium 1640 (1X), liquid RPMI Medium 1640 (1X), liquid Contains HEPES buffer Contains no ribonucleosides or deoxyribonucleosides Contains ribonucleosides and deoxyribonucleosides Contains Earles salts Contains Earles salts and HEPES buffer Low glucose Contains sodium pyruvate High glucose Contains sodium pyruvate High glucose Contains no sodium pyruvate High glucose Contains HEPES buffer but no sodium pyruvate Quantity 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) Cat. no. 21885-025 21885-108 31966-021 31966-047 61965-026 61965-059 32430-027 32430-100 31331-028 31331-093 31765-027 31765-068 31980-022 31980-048 32561-029 32561-094 32571-028 32571-093 41090-028 41090-093 42360-024 42360-081 51985-026 51985-042 61870-010 61870-044 72400-021 72400-054 Page 70 68 68 68 68 72 74 79 79 79 74 81 83 83
For stability data of GlutaMAX -I vs. L-glutamine, see Appendix I, page 202.
26
HEPES Media at a Glance

HEPES-buffered media resist fluctuations in pH caused by changes in cellular metabolism and culture environment. They are recommended for stabilizing the pH of sensitive cell culture systems. For your convenience, this page consolidates all gibco HEPES media in a concise, easy-to-use table. You can quickly scan it to determine what you need and where the product is fully described in this catalogue.
HEPES Media
Product Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid (high glucose) Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid (high glucose) Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid (low glucose) Dulbeccos Modified Eagle Medium (DMEM) (1X), liquid (high glucose) DMEM/F-12 (1X), liquid, 1:1 DMEM/F-12 (1X), liquid, 1:1 DMEM/F-12, powder, 1:1 F-10 Nutrient Mixture (Ham) (1X), liquid Iscoves Modified Dulbeccos Medium (IMDM) (1X), liquid Iscoves Modified Dulbeccos Medium (IMDM) (1X), liquid Iscoves Modified Dulbeccos Medium (IMDM) (1X), liquid Iscoves Modified Dulbeccos Medium (IMDM), powder McCoys 5A Medium (modified) (1X), liquid Medium 199 (1X), liquid Medium 199 (1X), liquid Minimum Essential Medium (MEM) (1X), liquid Minimum Essential Medium (1X MEM), liquid RPMI Medium 1640 (1X), liquid Contains 4,500 mg/L D-glucose, L-glutamine, and 25 mM HEPES buffer, but no sodium pyruvate Contains 4,500 mg/L D-glucose, L-glutamine, and 25 mM HEPES buffer, but no sodium pyruvate or phenol red Contains 1,000 mg/L D-glucose, L-glutamine, 25 mM HEPES buffer, and 110 mg/L sodium pyruvate Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine and 25 mM HEPES buffer, but no sodium pyruvate Contains 15 mM HEPES buffer and L-glutamine Contains 15 mM HEPES buffer and L-glutamine, but no phenol red Contains 15 mM HEPES buffer and L-glutamine, but no sodium bicarbonate Contains 25 mM HEPES buffer and L-glutamine Contains L-glutamine, 25 mM HEPES buffer, and 3,024 mg/L sodium bicarbonate, but no -thioglycerol or 2-mercaptoethanol Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, 25 mM HEPES buffer, and 3,024 mg/L sodium bicarbonate Contains L-glutamine, 25 mM HEPES buffer, and 3,024 mg/L sodium bicarbonate, but no -thioglycerol, 2-mercaptoethanol, or phenol red Contains L-glutamine and 25 mM HEPES buffer, but no sodium bicarbonate, -thioglycerol, or 2-mercaptoethanol Contains L-glutamine and 25 mM HEPES buffer Contains Earles salts, 2,200 mg/L sodium bicarbonate, L-glutamine, and 25 mM HEPES buffer Contains Hanks salts, 25 mM HEPES buffer, and L-glutamine Contains Earles salts and 25 mM HEPES buffer, but no L-glutamine Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, Earles salts, and 25 mM HEPES buffer Contains 25 mM HEPES buffer and L-glutamine Quantity 500 ml 10 500 ml (case) 500 ml 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 1L 1 10 L 500 ml 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 1 L 1 10 L 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 100 ml 500 ml 10 500 ml (case) 20 100 ml (case) 100 ml 500 ml 10 500 ml (case) 20 100 ml (case) 1L 1 10 L Cat. no. 42430-025 42430-082 21063-029 22320-022 22320-030 32430-027 32430-100 31330-038 31330-095 11039-021 11039-047 42400-010 42400-028 22390-025 21980-032 21980-065 31980-022 31980-048 21056-023 42200-014 42200-030 22330-021 22330-070 22340-020 22340-087 22350-029 22350-078 32360-026 32360-034 42360-024 42360-081 52400-017 52400-025 52400-041 52400-058 72400-013 72400-021 72400-054 72400-112 13018-015 13018-031 Page 68 69
69
Basal Media
70 71 71 71 72 74 74 74 74 75 76 76 79 79
83
RPMI Medium 1640 (1X), liquid
Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine and 25 mM HEPES buffer
83
RPMI Medium 1640, powder
Contains L-glutamine and 25 mM HEPES buffer, but no sodium bicarbonate
83
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Phenol Redfree Media at a Glance

Ideal for cell labeling experiments, phenol redfree media allow specific applications not possible with classical media. Phenol red, which is used as a pH indicator, can exhibit estrogen-like properties in some cells, and may be toxic to certain cell types. Gibco phenol redfree media are nontoxic and exhibit no estrogen-like activity in cell experimentation. For your convenience, this page consolidates all gibco phenol redfree media in a concise, easy-to-use table. You can quickly scan it to determine what you need and where the product is fully described in this catalogue.
Phenol Redfree Media

Product DMEM (1X), liquid (low glucose) DMEM (1X), liquid (high glucose) DMEM (1X), liquid (high glucose) DMEM/F-12 (1X), liquid, 1:1 DMEM/F-12 (1X), liquid, 1:1 Iscoves Modified Dulbeccos Media (IMDM) (1X), liquid Improved MEM Zn++ Option M (1X) (Richters Modification), liquid Leibovitzs L-15 Medium (1X), liquid Medium 199 (1X), liquid MEM Medium (1X), liquid MEM (1X), liquid Modified Eagle Medium (MEM) (2X), liquid Opti-MEM I Reduced-Serum Medium (1X), liquid RPMI Medium 1640 (1X), liquid Contains L-glutamine Contains L-glutamine Contains Earles salts, L-glutamine, and sodium bicarbonate Contains L-glutamine, ribonucleosides, and deoxyribonucleosides Contains Earles salts, but no L-glutamine Contains 1.0 g/L D-glucose and 110 mg/L sodium pyruvate (without L-glutamine) Contains 4.5 g/L D-glucose, L-glutamine, and 25 mM HEPES buffer Contains 4.5 g/L D-glucose (without L-glutamine) Contains L-glutamine and 15 mM HEPES buffer Contains L-glutamine Contains L-glutamine Quantity 500 ml 10 500 ml (case) 500 ml 500 ml 10 500 ml (case) 500 ml 10 500 ml (case) 500 ml 500 ml 500 ml 500 ml 500 ml 500 ml 500 ml 500 ml 500 ml 500 ml Cat. no. 11880-028 11880-036 21063-029 31053-028 11960-085 11039-021 11039-047 21041-025 21056-023 10373-017 21083-027 11043-023 41061-029 51200-038 21935-028 11058-021 11835-063 Page 70 68 68 71 71 74 74 75 76 78 79 80 81 83
Basal Medium Eagle (BME)

Basal Medium Eagle is well suited for use with diploid or primary mammalian cell cultures. For product formulation please refer to the tables at the end of this chapter.
BGJb Medium (Fitton-Jackson Modification)

BGJb Medium was formulated for the growth of fetal rat long bones in a chemically defined medium. For product formulation please refer to the tables at the end of this chapter.
Basal Medium Eagle (BME) (1X), liquid

Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD).
BGJb Medium (Fitton-Jackson Modification) (1X), liquid

Cat. no. 41010-026
Quantity 500 ml
Contains Earles salts, but no L-glutamine.
Quantity 500 ml
Contains L-glutamine.
Cat. no. 12591-038
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Brinsters BMOC-3 Medium

Brinsters BMOC-3 Medium, liquid
For product formulation please refer to the tables at the end of this chapter. Recommended storage conditions: 2C to 8C.
CMRL Medium
CMRL Medium is well suited for growth of Earles L cells. It is especially useful for cloning monkey kidney cell cultures and for growth of other mammalian cell lines when enriched with horse or calf serum. For product formulation please refer to the tables at the end of this chapter.
Quantity 100 ml
Contains bovine serum albumin, but no antibiotics.
CMRL Medium, 1066 (1X), liquid

Cat. no. 11126-034
3
Quantity Cat. no. 21530-027 500 ml
Contains no L-glutamine.
Cryopreservation Medium
Recovery Freezing Medium is suitable for the consistent cryopreservation of a broad spectrum of mammalian cells.
Basal Media
CMRL Medium, 1066 (10X), liquid

Recovery Freezing Medium

Recommended storage conditions: 5C to 20C.
Quantity 500 ml Cat. no. 12648-010
Quantity 50 ml
Contains DMEM, fetal bovine serum, calf serum, and 10% dimethylsulfoxide (DMSO).
Cat. no. 21540-026
CO2-Independent Medium
This medium contains a buffering system that maintains a stable pH for extended periods of time under atmospheric CO2.
CO2-Independent Medium (1X), liquid
A non-HEPES proprietary medium suitable for supporting cell growth for a variety of epithelial, fibroblast, and lymphoid cell lines in atmospheric conditions.1 It is ideal for transporting cells or tissue and for the handling of mouse embryos in atmospheric conditions. Not intended for CO2-based mouse embryo culture. Recommended storage conditions: 2C to 8C, in the dark.
Reference
1. Battista, P.J. and Weiss, S.A. (1991) FOCUS 13, 110.
Quantity 500 ml
Cat. no. 18045-054
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Dulbeccos Modified Eagle Media (DMEM)

Dulbeccos Modified Eagle Media are well suited for supporting the growth of a broad spectrum of mammalian cell lines. Use the comparison tables as quick references for identifying the catalogue number of the product thats best for your application.
Advanced DMEM (1X), liquid

Recommended storage conditions: 2C to 8C, in the dark.
Quantity 500 ml 10 500 ml (case) Cat. no. 12491-015 12491-023
Reference
Price, P. and Staines, D. (2004) Advanced Media: Reduced Sera Solutions, Quest 3, 2530.
Contains 4,500 mg/L D-glucose, non-essential amino acids (NEAA), and 110 mg/L of sodium pyruvate, but no L-glutamine.
DMEM (1X), liquid (high glucose)

For product formulation please refer to the tables at the end of this chapter. Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD).
Quantity 500 ml 10 500 ml (case) 1,000 ml 6 1,000 ml (case) Cat. no. 41965-039 41965-062 41965-047 41965-120 Quantity 1 10 L Cat. no. 52100-039
Contains 4,500 mg/L D-glucose and L-glutamine, but no sodium pyruvate or sodium bicarbonate.
Contains 4,500 mg/L D-glucose and L-glutamine, but no sodium pyruvate.
Quantity 1 10 L
Cat. no. 12800-116
Quantity 500 ml 10 500 ml (case)
Cat. no. 41966-029 41966-052
Contains 4,500 mg/L D-glucose, L-glutamine, and 110 mg/L sodium pyruvate, but no sodium bicarbonate.
Cat. no. 11960-044 11960-085
Contains 4,500 mg/L D-glucose, L-glutamine, and 110 mg/L sodium pyruvate.
Cat. no. 42430-025 42430-082
Contains 4,500 mg/L D-glucose, but no L-glutamine or sodium pyruvate.
Quantity 500 ml
Cat. no. 21969-035
Contains 4,500 mg/L D-glucose, L-glutamine, and 25 mM HEPES buffer, but no sodium pyruvate.
Quantity 500 ml
Cat. no. 21063-029
Contains 4,500 mg/L D-glucose and 110 mg/L sodium pyruvate, but no L-glutamine.
Quantity 500 ml
Cat. no. 31053-028
Contains 4,500 mg/L D-glucose, L-glutamine, and 25 mM HEPES buffer, but no sodium pyruvate or phenol red.
Contains 4,500 mg/L D-glucose, but no L-glutamine, sodium pyruvate, or phenol red.
Quantity 500 ml
Cat. no. 11971-025
Quantity 500 ml
Cat. no. 21013-024
Contains 4,500 mg/L D-glucose and L-glutamine, but no sodium phosphate or sodium pyruvate.
Contains 4,500 mg/L D-glucose, but no L-glutamine, sodium pyruvate, L-methionine, or L-cystine.
30
Quantity 500 ml
Cat. no 21068-028
Cat. no. 61965-026 61965-059
Contains 4,500 mg/L D-glucose, but no L-glutamine, sodium pyruvate, or calcium chloride.
Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine and 4,500 mg/L D-glucose, but no sodium pyruvate.
Cat. no. 31966-021 31966-047
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, 4,500 mg/L D-glucose, and 110 mg/L sodium pyruvate.
Cat. no. 32430-027 32430-100
Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, 4,500 mg/L D-glucose, and 25 mM HEPES buffer, but no sodium pyruvate.
3 Basal Media
Comparison of Dulbeccos Modified Eagle Media (DMEM), High Glucose (4,500 mg/L)
Base cat. no. With L-glutamine With Na+ pyruvate (110 mg/L) With HEPES buffer (25 mM) Without phenol red Without Na+ phosphate Powder
41965*
41966
42430
21063
11971
52100*
12800
11960* 21969 31053 32430 61965 31966
21068 12491
Without L-glutamine With GlutaMAX-I With HEPES buffer (25 mM) With Na pyruvate (110 mg/L) Without phenol red Without CaCl2 Without cystine Without methionine
+
* Standard formulation, as published in Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J.D., Freeman, G., Vogt, M., and Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 6:2, 93. Powdered formulations do not contain sodium bicarbonate. GlutaMAX-I supplement is L-alanyl-L-glutamine, a glutamine dipeptide, which can be used in place of L-glutamine in culture media. It is substituted on an equimolar basis for L-glutamine. Use this comparison table as a quick reference for identifying the base catalogue number of the product thats best for your application. For the exact composition of each product, please refer to the formulation tables starting on page 88.
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DMEM (1X), liquid (low glucose)
For product formulation please refer to the tables at the end of this chapter. Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD).
DMEM (1X), liquid (no glucose)

Contains L-glutamine, but no glucose or sodium pyruvate.
Contains 1,000 mg/L D-glucose, L-glutamine, and 110 mg/L sodium pyruvate.
DMEM, powder (low glucose)

Quantity 500 ml
Cat. no. 22320-022
Contains 1,000 mg/L D-glucose, L-glutamine, 25 mM HEPES buffer, and 110 mg/L sodium pyruvate.
Quantity 1 10 L 1 50 L Cat. no. 31600-083 31600-091
Quantity 500 ml
Cat. no. 11880-028
Contains 1,000 mg/L D-glucose and 110 mg/L sodium pyruvate, but no L-glutamine or phenol red.
Contains 1,000 mg/L D-glucose, L-glutamine, and 110 mg/L sodium pyruvate, but no sodium bicarbonate.
Intended use: In vitro diagnostic (IVD).

A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, 1,000 mg/L D-glucose, and 110 mg/L sodium pyruvate.
Comparison of Dulbeccos Modified Eagle Media (DMEM), low glucose (1,000 mg/L)
Base cat. no. With L-glutamine With HEPES buffer (25 mM) Without glucose Powder Without L-glutamine With GlutaMAX-I Without phenol red 11880 21885 31885* 11966 22320 31600*
* Standard formulation, as published in Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J.D., Freeman, G., Vogt, M., and Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 6:2, 93. Powdered formulations do not contain sodium bicarbonate. GlutaMAX-I supplement is L-alanyl-L-glutamine, a glutamine dipeptide, which can be used in the place of L-glutamine in culture media. It is substituted on an equimolar basis for L-glutamine. Use this comparison table as a quick reference for identifying the base catalogue number of the product thats best for your application. For the exact composition of each product, please refer to the formulation tables starting on page 88.
32
DMEM Nutrient Mixture F-12 (DMEM/F-12) 1:1

Advanced DMEM/F-12 (1X), liquid, 1:1
Recommended storage conditions: 2 to 8C, in the dark.
Reference
Price, P. and Staines, D. (2004) Advanced Media: Reduced Sera Solutions, Quest 3, 2530. www.invitrogen.com/advanced
Contains 3,151 mg/L D-glucose, non-essential amino acids (NEAA), and 110 mg/L sodium pyruvate, but no L-glutamine.
Cat. no. 12634-010 12634-028
DMEM/F-12 (1X), liquid, 1:1

Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD). For product formulation please refer to the tables at the end of this chapter.
Basal Media
Quantity 500 ml 10 500 ml (case) 10 L
Contains 15 mM HEPES buffer and L-glutamine.
Cat. no. 31330-038 31330-095 11330-107

Contains 15 mM HEPES buffer and L-glutamine, but no phenol red.
Cat. no. 11039-021 11039-047
Quantity 500 ml
Contains L-glutamine, but no HEPES buffer or phenol red.
Cat. no. 21041-025
Cat. no. 31331-028 31331-093
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, but no HEPES buffer.
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F-10 Nutrient Mixtures

F-10 Nutrient Mixture (Ham) was formulated for serum-free growth of Chinese hamster ovary (CHO) cells and growth of various mammalian cell lines with serum supplementation. For product formulation please refer to the tables at the end of this chapter.

F-12 Nutrient Mixture was formulated for single-cell plating of near-diploid Chinese hamster ovary (CHO) cells.
F-12 Nutrient Mixture (Ham) (1X), liquid

F-10 Nutrient Mixture (Ham) (1X), liquid

Quantity 500 ml
Quantity Cat. no. 31550-023 500 ml 10 500 ml (case) 1,000 ml Cat. no. 22390-025
Cat. no. 21765-029 21765-037 11765-047
Quantity 500 ml
Cat. no. 31765-027 31765-068
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine.
F-12 Nutrient Mixture (Ham), powder

Quantity 10 1 L 1 50 L
Contains L-glutamine, but no sodium bicarbonate.
Cat. no. 21700-026 21700-018
F-12K Nutrient Mixture, Kaighns Modification (1X), liquid

Recommended storage conditions: 2C to 8C. For product formulation please refer to the tables at the end of this chapter.
Quantity 500 ml
Cat. no. 21127-022
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Glasgow Minimum Essential Medium

Glasgow Minimum Essential Medium was formulated to support the growth of BHK-21 cells. For product formulation please refer to the tables at the end of this chapter.
Graces Insect Cell Culture Media

For product formulation please refer to the tables at the end of this chapter.
Glasgow Minimum Essential Medium (G-MEM, BHK-21) (1X), liquid

Graces Insect Cell Culture Medium, supplemented (1X), liquid

Insect cell culture tested. Recommended storage conditions: 2C to 8C, in the dark.
Quantity 500 ml Cat. no. 21710-025 10 500 ml (case) Cat. no. 11605-045 11605-086
Quantity 500 ml
Contains L-glutamine, but no tryptose phosphate broth.
Contains 3330 mg/L lactalbumin hydrolysate, 3330 mg/L yeastolate, and L-glutamine, but no insect hemolymph.
Basal Media

Quantity 100 ml
Contains lactalbumin hydrolysate, yeastolate, and L-glutamine.
Cat. no. 11667-037
Graces Insect Cell Culture Medium, unsupplemented (1X), liquid

Quantity 500 ml
Contains L-glutamine, but no L-methionine or insect hemolymph.
Cat. no. 11595-030
Graces Insect Cell Culture Medium, powder

Quantity 10 1 L
Cat. no. 11300-027
Related products Page Insect cell expression products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98 Sf9 cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175
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Improved MEM Zn++ Option (Richters Modification)

Iscoves Modified Dulbeccos Media

Iscoves Modified Dulbeccos Media are highly enriched synthetic media. They are well suited for rapidly proliferating, high-density cell cultures. For product formulation please refer to the tables at the end of this chapter.
Improved MEM Zn++ Option (Richters Modification), liquid

Iscoves Modified Dulbeccos Medium (IMDM) (1X), liquid

Cat. no. 10373-017
Quantity 500 ml
Contains L-glutamine, 2 mg/L L-proline, and 50 g/ml gentamicin sulfate, but no insulin, HEPES buffer, or phenol red.
Quantity 500 ml 10 500 ml (case) 1,000 ml Cat. no. 21980-032 21980-065 12440-046
Contains L-glutamine, 25 mM HEPES buffer, and 3,024 mg/L sodium bicarbonate, but no -thioglycerol or 2-mercaptoethanol.
IPL-41 Insect Medium

Cat. no. 31980-022 31980-048
IPL-41 Insect Medium (1X), liquid

A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, 25 mM HEPES buffer, and 3,024 mg/L sodium bicarbonate.
Quantity 500 ml
Cat. no. 21056-023
Contains L-glutamine, calcium chloride, and sodium bicarbonate, but no tryptose phosphate broth.
Contains L-glutamine, 25 mM HEPES buffer, and 3,024 mg/L sodium bicarbonate, but no -thioglycerol, 2-mercaptoethanol, or phenol red.
Iscoves Modified Dulbeccos Medium (IMDM), powder

Quantity 1L 10 L Cat. no. 42200-014 42200-030
Contains L-glutamine and 25 mM HEPES buffer, but no sodium bicarbonate, -thioglycerol, or 2-mercaptoethanol.
Related products Page Insect cell expression products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
36
Leibovitzs L-15 Media

Leibovitzs L-15 Media are suited for supporting cell growth in nonCO2-equilibrated environments. These formulations were developed without a sodium bicarbonate buffer system. For product formulation please refer to the tables at the end of this chapter.
McCoys 5A Medium (Modified)

McCoys Medium (modified) was formulated for the propagation of human lymphocytes. It was based on the original formulation of McCoy as modified by Hsu and Kellog and further modified by Iwakata and Grace. For product formulation please refer to the tables at the end of this chapter.
Leibovitzs L-15 Medium (1X), liquid

McCoys 5A Medium (modified) (1X), liquid

3
Cat. no. 11415-049 11415-056 Quantity 500 ml 10 500 ml (case)
Cat. no. 26600-023 26600-080
Basal Media

Quantity 500 ml
Contains L-glutamine, but no phenol red.

Contains L-glutamine and 25 mM HEPES buffer.
Cat. no. 22330-021
Leibovitzs L-15 Medium, powder

MCDB 131 Medium

Cat. no. 41300-021 41300-070
Quantity 1L 10 L
MCDB 131 was developed by Knedler and Ham as a reduced serumsupplemented medium for the culture of human microvascular endothelial cells. This formulation has been reported to support clonal growth of human omental microvascular cells when supplemented with 0.7% dialyzed FBS, 10 ng/ml EGF, and 1 g/ml hydrocortisone. For product formulation please refer to the tables at the end of this chapter.
Reference
Knedler and Ham (1987) In Vitro Cell. Develop. Biol. 23, 481.
MCDB 131 Medium (1X), liquid

Quantity 500 ml
Cat. no. 10372-019
37
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Media 199
Media 199 were formulated for nutritional studies of chick embryo fibroblasts. Media 199 are formulated with either Earles salts or Hanks salts. Media utilizing Earles salts are buffered with a bicarbonate/carbonic acid system and will maintain pH in a CO2 incubator. Use of Earles salts in atmospheric conditions will result in rapid rise in pH of the culture medium. Media utilizing Hanks salts are buffered with salt solutions designed for atmospheric equilibration, and use in a CO2 incubator will result in a rapid drop in pH of the culture medium. For product formulation please refer to the tables at the end of this chapter.
Medium 199 with Earles Salts

Medium 199 (1X), liquid
Medium 199, powder

Quantity 1L Cat. no. 31150-022 31150-030 1 50 L
Contains Earles salts and L-glutamine, but no sodium bicarbonate.
Contains L-glutamine and 2,200 mg/L sodium bicarbonate.
Cat. no. 31100-027 31100-019
Medium 199 with Hanks Salts

Cat. no. 22340-020
Quantity 500 ml

Contains 2,200 mg/L sodium bicarbonate, L-glutamine, and 25 mM HEPES buffer.
Quantity 500 ml
Cat. no. 11043-023
Quantity 500 ml
Contains L-glutamine and 2,200 mg/L sodium bicarbonate, but no phenol red.
Cat. no. 22350-029

Quantity 500 ml
Contains L-glutamine but no sodium bicarbonate.
Cat. no. 21180-021
38
Minimum Essential Media (MEM)

Minimum Essential Media are patterned after Eagles Media and are well suited for the growth of a broad spectrum of mammalian cells.
Comparison of Minimum Essential Media (MEM) with Earles Salts

Base cat. no. With L-glutamine With non-essential amino acids With ribonucleosides and deoxyribonucleosides Without phenol red -modification Powder Without L-glutamine With HEPES buffer (25 mM) With non-essential amino acids With GlutaMAX-I With ribonucleosides and deoxyribonucleosides Without phenol red -modification S-MEM modified for suspension culture (Ca free) 10X powder
* Standard formulation, as published in Eagle, H. (1959) Science 130, 432. MEM- glutamine dipeptide, which can be used in the place of L-glutamine in culture media. It is substituted on an equimolar basis for L-glutamine. Powdered media and concentrated formulations do not contain sodium bicarbonate. GlutaMAX-I supplement is L-alanyl-L-glutamine, a glutamine dipeptide, which can be used in place of L-glutamine in culture media. It is substituted on an equimolar basis for L-glutamine. Use this comparison table as a quick reference for identifying the base catalogue number of the product thats best for your application. For the exact composition of each product, please refer to the formulation tables starting on page 100.
2+
31095*
22571
22561
41061
61100*
41500
11900
12000
41090
32561
32571 21430 11700
21090*
10370
11380
32360
51200
Basal Media
39
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Advanced Minimum Essential Medium (MEM) (1X), liquid (with L-glutamine)
Recommended storage conditions: 2C to 8C, in the dark. For product formulation please refer to the tables at the end of this chapter. Additional information is available at www.invitrogen.com/advanced.
Contains L-glutamine, but no ribonucleosides or deoxyribonucleosides.
Quantity 500 ml
Cat. no. 41061-029
Reference
Contains L-glutamine, ribonucleosides, and deoxyribonucleosides, but no phenol red.
Minimum Essential Medium (MEM), powder

Contains 110 mg/L of sodium pyruvate, non-essential amino acids (NEAA), but no L-glutamine.
Quantity Cat. no. 61100-087 61100-103 1 10 L 1 50 L
Contains Earles salts and L-glutamine, but no sodium bicarbonate.
Minimum Essential Medium (MEM) with Earles Salts (with L-glutamine)

Minimum Essential Medium (MEM) (1X), liquid
Quantity 1 10 L 1 50 L
Cat. no. 41500-018 41500-083
Contains Earles salts, L-glutamine, and non-essential amino acids, but no sodium bicarbonate.
Contains Earles salts and L-glutamine.
Cat. no. 31095-029 31095-052
Minimum Essential Medium (MEM) Medium, powder

Minimum Essential Medium (MEM) Medium (1X), liquid

Quantity 1 10 L Cat. no. 11900-073
Contains L-glutamine, ribonucleosides, and deoxyribonucleosides, but no sodium bicarbonate.
Contains L-glutamine, ribonucleosides, and deoxyribonucleosides.
Quantity Cat. no. 22571-020 22571-038 1 10 L
Cat. no. 12000-063
Contains L-glutamine, but no ribonucleosides, deoxyribonucleosides, or sodium bicarbonate.
40
Minimum Essential Medium (MEM) with Earles Salts (without L-glutamine)

Minimum Essential Medium (MEM) Medium (1X), liquid

Cat. no. 21090-022 21090-055
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, but no ribonucleosides or deoxyribonucleosides.
Quantity Cat. no. 10370-047 10370-070 500 ml 10 500 ml (case)
Cat. no. 32571-028 32571-093

Contains non-essential amino acids, but no L-glutamine.
Basal Media
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, ribonucleosides, and deoxyribonucleosides.
Quantity 500 ml
Contains 25 mM HEPES buffer, but no L-glutamine.
Cat. no. 32360-026
Quantity 500 ml
Contains no L-glutamine or phenol red.
Cat. no. 51200-046
Cat. no. 41090-028 41090-093
Cat. no. 42360-024 42360-081
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine and 25 mM HEPES buffer.

Quantity 500 ml
Contains no L-glutamine or sodium bicarbonate.
Cat. no. 21430-020
41
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Minimum Essential Medium (MEM) with Hanks' Salts, liquid (with L-glutamine)

F-10 Nutrient Mixture (Ham) was formulated for serum-free growth of Chinese hamster ovary (CHO) cells and growth of various mammalian cell lines with serum supplementation. For ordering information, please refer to page 34.

F-12 Nutrient Mixture was formulated for single-cell plating of neardiploid Chinese hamster ovary (CHO) cells. For ordering information, please refer to page 34.
Modified Eagle Media

Modified Eagle Medium (MEM) (2X), liquid
Medium NCTC-109
NCTC-109 is a synthetic medium that is suitable for general use in generating and maintaining hybridoma cells. This medium typically requires the addition of a growth factor cocktail or serum. For product formulation please refer to the tables at the end of this chapter.
Medium NCTC-109 (1X), liquid

Quantity 500 ml
Cat. no. 21340-054
Related products Page Hybridoma and monoclonal antibody products. . . . . . . . . . . . . . . . . . . . .106
42
Opti-MEM I Reduced-Serum Media

Opti-MEM I is a modification of Eagles Minimum Essential Medium, buffered with HEPES and sodium bicarbonate (2.4 g/L) and supplemented with hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, and growth factors. The concentration of phenol red has been reduced to 1.1 mg/L. Most cells routinely cultured in serum-supplemented medium may be transferred directly into Opti-MEM I with a minimum of 50% reduction in serum. Further significant reductions in serum requirements have been achieved with myelomas (Sp2/O-Ag14, P363 Ag8.653, P3-NSI-Ag4-1) and derived hybridomas, as well as with fibroblasts and epithelial cells of both normal and tumor origins. For very low serum supplementation (<1%) with attachment-dependent cells, add 500 to 1,000 mg/L calcium chloride. Ideal for use during cationic lipid transfections.1
Reference
1. FOCUS on Applications, Opti-MEM I Reduced-Serum Medium: Application in Transfection, Technical Bulletin.
Opti-MEM I Reduced-Serum Medium (1X), liquid

Recommended storage conditions: 2C to 8C, in the dark. Intended use: In vitro diagnostic (IVD).

Recommended storage conditions: 2C to 8C, in the dark. Intended use: In vitro diagnostic (IVD). Note: See page 84 for procedure to prepare powdered Opti-MEM I.
3 Basal Media
Quantity 100 ml 500 ml 10 500 ml (case) Cat. no. 31985-062 31985-047 31985-054
A modification of MEM (Eagles). Contains HEPES buffer, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1.1 mg/L, but no sodium bicarbonate.
Contains HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1.1 mg/L.
Opti-MEM I Reduced-Serum Medium (1X), liquid (without phenol red)

Lipofectamine 2000 Transfection Reagent

Lipofectamine 2000 Transfection Reagent is a proprietary cationic lipid formulation that offers the highest transfection efficiencies on the widest variety of cell lines with high protein expression levels or gene knockdown activity (RNAi). It is the leading transfection reagent for simple, effective, and optimal nucleic acid delivery in a wide range of eukaryotic cell types and is proven to work in high-throughput applications. Recommended storage conditions: 2C to 8C.
Contains HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, and growth factors, but no phenol red.
Quantity 0.75 ml 1.5 ml Cat. no. 11668-027 11668-019
Opti-MEM I Reduced-Serum Medium (1X), liquid (with GlutaMAX-I)

Recommended storage conditions: 2C to 8C, in the dark. Intended use: In vitro diagnostic (IVD).
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, trace elements, growth factors, and phenol red reduced to 1.1 mg/L.
43
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RPMI Media 1640

RPMI Media 1640 are enriched formulations with extensive applications for mammalian cells. They were formulated for suspension cultures or monolayer culture of human leukemic cells.
RPMI Media 1640 Comparison Chart

Base cat. no. With L-glutamine With HEPES buffer (25 mM) Without glucose Without phenol red Without folate Powder
21875*
11835
11879
52400
51800*
13018
27016
31870* 61870 72400
Without L-glutamine With GlutaMAX-I With HEPES buffer (25 mM)
* Standard formulation, as published in Moore, G.E., Gerner, R.E., and Franklin, H.A. (1967) JAMA 199, 519. Powdered media do not contain sodium bicarbonate. GlutaMAX-I supplement is L-alanyl-L-glutamine, a glutamine dipeptide, which can be used in the place of L-glutamine in culture media. It is substituted on an equimolar basis for L-glutamine. Use this comparison table as a quick reference for identifying the base catalogue number of the product thats best for your application. For the exact composition of each product, please refer to the formulation tables on page 107.
44
Advanced RPMI Medium 1640 (1X), liquid

Recommended storage conditions: 2C to 8C, dark
www.invitrogen.com/advanced
Cat. no. 61870-010 61870-044
Reference
Cat. no. 72400-021 72400-054
A GlutaMAX-I medium. Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine and 25 mM HEPES buffer.
Contains 2,000 mg/L D-glucose, non-essential amino acids (NEAA), and 110 mg/L sodium pyruvate, but no L-glutamine.
RPMI Medium 1640, powder

RPMI Medium 1640 (1X), liquid

Basal Media
Quantity 1L 1 10 L Cat. no. 51800-019 51800-035 51800-043
Quantity 100 ml 500 ml 10 500 ml (case) 6 1,000 ml (case)
Cat. no. 21875-042 21875-034 21875-091 21875-158
1 50 L
Quantity 1L 1 10 L
Cat. no. 13018-015 13018-031
Contains L-glutamine and 25 mM HEPES buffer, but no sodium bicarbonate.
Quantity 500 ml
Contains L-glutamine, but no phenol red.
Cat. no. 11835-063
Quantity 100 ml 500 ml 10 500 ml (case)

Contains L-glutamine and 25 mM HEPES buffer.
Cat. no. 52400-017 52400-025 52400-041
Quantity 500 ml
Contains L-glutamine, but no glucose.
Cat. no. 11879-020
Quantity 500 ml
Contains L-glutamine, but no folic acid.
Cat. no. 27016-021

Cat. no. 31870-025 31870-074
45
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Schneiders Drosophila Medium

Williams Medium E
Williams Medium E was formulated for long-term cell cultures of adult rat liver epithelial cells. For product formulation please refer to the tables at the end of this chapter.
Schneiders Drosophila Medium (1X), liquid

Modified and insect cell culture tested. Recommended storage conditions: 2C to 8C.
Williams Medium E (1X), liquid

Cat. no. 21720-024
Quantity 500 ml
Quantity 500 ml
Cat. no. 22551-022
Waymouths MB 752/1 Medium

Waymouths MB 752/1 Media was formulated for studies on the nutrition, metabolism, and growth of Strain L sublines, NCTC clone 929. For product formulation please refer to the tables at the end of this chapter.
Waymouths MB 752/1 Medium (1X), liquid

Quantity 500 ml
Cat. no. 31220-023
46
Media Preparation
Liquid Media: Preparing 1X Solutions from 10X Concentrates
To prepare an acceptable final 1X solution, perform the following procedures under aseptic conditions. We recommend distilled water (base cat. no. 15230) and 7.5% Sodium Bicarbonate Solution (base cat. no. 25080) for use in this protocol. Use sterile solutions of 1 N sodium hydroxide or hydrochloric acid prepared by membrane filtration using appropriate filters. Also note that because we adjust the pH of all 10X media and 10X balanced salt solution concentrates for solubility, you may need to adjust the pH after dilution and add sodium bicarbonate, as appropriate. 1. Aseptically dilute 100 ml of 10X concentrate with approximately 850 ml of distilled water. 2. Aseptically add the correct amount of 7.5% sodium bicarbonate solution (see table below for concentrations). 3. Adjust the pH as necessary with 1 N hydrochloric acid or 1 N sodium hydroxide (see table below for 1X pH values). 4. QS to volume with distilled water. 5. Dispense the solution into sterile containers. Cap the bottles tightly with sterile closures and store them at the recommended temperature (see product descriptions in this chapter).
3 Basal Media
Procedure 1: Preparing powdered media

1. To a mixing container that is as close to the final volume as possible, add 23% less distilled water than the desired total volume of medium. For example, for 1 L of medium start with 970 ml to 980 ml of distilled water. 2. Add powdered medium to room temperature (20C to 30C) water with gentle stirring. Do not heat the water. 3. Rinse the inside of the package to remove all traces of powder. 4. Add NaHCO3 (see table below for quantity) to medium. See the table on the next page for sodium bicarbonate concentration. 5. Adjust the pH to 0.2 and 0.3 units below the desired final working pH by slowly adding, with stirring, either 1 N NaOH or 1 N HCl. The pH may rise 0.1 to 0.3 units upon filtration. 6. Dilute the medium to the desired volume with distilled water. Do not overmix. Keep the container closed until the medium is filtered. 7. Process the medium immediately into sterile containers by membrane filtration using a 0.2 m filter. We recommend using a positive-pressure system.
Procedure 2: Preparing powdered Leibovitzs L-15 Media

1. To a mixing container that is as close to the final volume as possible, add 23% less distilled water than the desired total volume of medium. 2. Add powdered medium to room temperature (20C to 30C) water with gentle stirring. Do not heat the water. 3. Rinse the inside of the package to remove all traces of powder. 4. Adjust the pH to 7.6 by slowly adding, with stirring, 1 N NaOH or 1 N HCl. 5. Dilute the liquid to the desired volume with distilled water and stir until dissolved. Do not overmix. 6. Process the medium immediately into sterile containers by membrane filtration using a 0.2 m filter. We recommend using a positive-pressure system.
Sodium Bicarbonate Concentrations Achieved Using 7.5% Sodium Bicarbonate Solution (base cat. no. 25080)
NaHCO3 pH of 10X solution Base cat. no. 14065 14080 14185 14200
Final concentration (g/L)
7.5% solution to add (ml/L)
pH range after 1:10 dilution and addition of NaHCO3
Recommended pH 1X solution*
Balanced salt solutions 5.86.1 4.44.7 5.86.1 6.77.0 0.35 0.35 4.7 4.7 Liquid media 7.57.8 5.25.5 6.06.3 7.17.4 7.07.4 7.07.2 7.07.4 7.07.2
21430
4.95.4
2.2
29.3
7.67.9
7.07.4
* pH range will vary depending on pH of water used for dilution. Formulation does not require addition of NaHCO3.
47
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Procedure 3: Preparing powdered Iscoves Modified Dulbeccos Media

1. To a mixing container that is as close to the final volume as possible, add 23% less distilled water than the desired total volume of medium. 2. Add powdered medium to room temperature (20C to 30C) water with gentle stirring. Do not heat the water. 3. Rinse the inside of the package to remove all traces of powder. 4. Add 3.024 g of NaHCO3/L of medium. 5. Dilute the medium to the desired volume with distilled water and stir until dissolved. Do not overmix. 6. Do not adjust the pH. Keep the container closed until the medium is filtered. 7. Process the medium immediately into sterile containers by membrane filtration using a 0.2 m filter. We recommend using a positivepressure system.
Procedure 4: Preparing powdered Opti-MEM I ReducedSerum Medium

1. To a mixing container that is as close to the final volume as possible, add 23% less distilled water than the desired total volume of medium. For example, for 1 L of medium start with 970 ml to 980 ml of distilled water. 2. Add powdered medium to room temperature (20C to 30C) water with gentle stirring. Do not heat the water. 3. Rinse the inside of the package to remove all traces of powder. 4. Stir until completely in solution (usually within 10 min). 5. Adjust the pH to between 0.2 and 0.3 units below the desired final pH by slowly adding, with stirring, either 1 N NaOH or 1 N HCl (recommended final pH is 7.3 0.1). The pH may rise 0.1 to 0.3 units upon filtration. 6. Add 2.4 g of reagent-grade NaHCO3 and, if appropriate, 1.0 ml of 5.5 102 M 2-mercaptoethanol (1,000X)/L of medium. 7. Dilute the medium to the desired volume with distilled water and stir until dissolved. Do not overmix. Keep the container closed until the medium is filtered. 8. Process the medium immediately into sterile containers by membrane filtration using a 0.2 m filter. We recommend using a positive-pressure system. Lowprotein-adsorbent filters are required.
Preparation of Powdered Media

NaHCO3 Procedure number* 1 1 1 1 3 1 1 1 1 1 4 5 1 1 5 2 1 1 g/L 0.35 2.2 2.2 3.7 3.02 (1.2) 1,125 2.438 3.7 0 1.18 2.4 2.0 2.2 3.7 2.0 2.2 2.2 ml/L 7.5% solution 4.7 29.3 29.3 49.3 40.3 (16.0) 15 (32.5) 15 49.3 0 15.7 32.0 26.7 29.3 49.3 26.7 29.3 29.3 Recommended working pH 6.06.2 7.07.4 7.07.4 7.07.4 (6.97.3) 7.07.4 7.17.4 7.17.4 7.07.4 7.07.2 7.67.9 7.27.4 7.07.4 (7.17.4) 7.07.4 7.07.4 7.07.4 7.37.6 7.07.4 7.07.4 31600 51800 41300 41500 61100 42400 32500 12800 21600 21700 22600 22340 31100 Base cat. no. 11300 11900 12000 52100 42200
Procedure 5: Preparing powdered RPMI 1640 Media

1. To a mixing container that is as close to the final volume as possible, add 23% less distilled water than the desired total volume of medium. For example, for 1 L of medium start with 970 ml to 980 ml of distilled water. 2. Add powdered medium to room temperature (20C to 30C) water with gentle stirring. Do not heat the water. 3. Rinse the inside of the package to remove all traces of powder. 4. Add 2.0 g of NaHCO3/L of medium (it will appear cloudy because of incomplete solubilization). 5. Adjust the pH to between 0.2 and 0.3 below the desired final working pH by slowly adding, with stirring, either 1 N NaOH or 1 N HCl. The pH may rise 0.1 to 0.3 units upon filtration. 6. Dilute the medium to the desired volume with distilled water and stir until dissolved. Do not overmix. Keep the container closed until the medium is filtered. 7. Process the medium immediately into sterile containers by membrane filtration using a 0.2 m filter. We recommend using a positive-pressure system.
* See procedures 1 through 5. Formulations do not require addition of NaHCO3.
48
E-Z Hold Plastic Bottles

GIBCO E-Z Hold Bottles were designed for cell culturists from polyethylene terephthalate (PET) plastic, the most recycled plastic in the world. These bottles are lightweight and virtually unbreakable. Bottle features include a wide mouth and no-drip spout for easy pipetting, two flat sides for easy gripping, and an overall round shape to facilitate swirling of contents in the bottle. This product is the subject of US Patent 5,246,127.
Stak Pack Accessories

Stak Pack accessories are for use with the GIBCO Stak Pack delivery system. The contents of each package are radiation sterilized and are suitable for aseptic cell culture procedures unless the package has been opened or damaged. This product is the subject of US Patent 4,869,398.
Product
36" transfer tube with male luer lock; radiation sterilized
Product
100 ml E-Z Hold bottles with caps 500 ml E-Z Hold bottles with caps 1,000 ml E-Z Hold bottles with caps
Quantity 1 1 1
Cat. no. 12002-010 10265-015 11003-019
Quantity 264 units/case 66 units/case 24 units/case
Cat. no. 10833-010 10833-028 10833-036
36" transfer tube with female luer lock; radiation sterilized 8" interconnecting tube; radiation sterilized
3 Basal Media 49
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Media Formulations
Basal Medium Eagle*
Base cat. no. Component Inorganic salts CaCl2 (anhyd.) KCl MgSO4 (anhyd.) NaCl NaHCO3 NaH2PO4H2O Other components D-Glucose Phenol red Amino acids L-Arginine HCl L-Cystine 2HCl L-Histidine L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Threonine L-Tryptophan L-Tyrosine 2Na 2H2O L-Valine Vitamins Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide Pyridoxal HCl Riboflavin Thiamine HCl 1.00 1.00 1.00 1.00 1.80 1.00 1.00 0.10 1.00 21.00 12.00 8.00 26.00 26.00 29.00 7.50 16.50 24.00 4.00 18.00 23.50 1000.00 10.00 264.00 400.00 200.00 6800.00 2200.00 158.00 41010 1X liquid, mg/L
CMRL 1066 Medium 1

Base cat. no. Component Inorganic salts CaCl2 (anhyd.) KCl MgSO4 .7H2O NaCl NaHCO3 NaH2PO4 H2O Other components Cocarboxylase Coenzyme A 2-Deoxyadenosine 2-Deoxycytidine HCl 2-Deoxyguanosine Diphosphopyridine nucleotide Ethanol for solubilsing lipid components Flavin adenine dinucleotide D-Glucose Glutathione (reduced) 5-Methyl-deoxycytidine Phenol red* Sodium acetate 3H2O Sodium glucuronate H2O Thymidine Triphosphopyridine nucleotide Tween 80 Uridine triphosphate Amino acids L-Alanine L-Arginine HCl L-Aspartic acid L-Cystine (free base) L-Cystine 2HCl L-Glutamic acid Glycine L-Histidine HCl H2O Hydroxy-L-proline L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine 25.00 70.00 30.00 199.88 20.00 75.00 50.00 20.00 10.00 20.00 60.00 70.00 15.00 25.00 40.00 25.00 250.00 700.00 300.00 1998.80 200.00 750.00 500.00 200.00 100.00 200.00 600.00 700.00 150.00 250.00 400.00 250.00 1.00 2.50 10.00 10.00 10.00 7.00 16.0 1.00 1000.00 10.00 0.10 20.00 83.00 4.00 10.00 1.00 5.00 1.00 10.00 25.00 100.00 100.00 100.00 70.00 160.00 10.00 10000.00 100.00 1.00 200.00 830.00 40.00 100.00 10.00 50.00 10.00 264.00 400.00 200.00 6799.00 2200.00 158.00 2000.00 4000.00 2000.00 67990.00 1580.00 21530 1X liquid, mg/L 21540 10X liquid, mg/L
L-Threonine L-Tryptophan L-Tyrosine L-Valine
30.00 10.00 40.00 25.00
300.00 100.00 400.00 25.00
CMRL 1066 Medium 1

Base cat. no. Component 1X liquid, mg/L 50.00 0.01 0.01 0.20 0.50 0.01 0.05 0.025 0.025 0.05 0.025 0.01 0.01 10X liquid mg/L 500.00 0.10 0.10 2.00 5.00 0.10 0.50 0.25 0.25 0.5 0.254 0.1 0.10
Vitamins Ascorbic acid D-Biotin D-Ca pantothenate Cholesterol Choline chloride Folic acid i-Inositol Niacin Niacinamide Para-aminobenzoic acid Pyridoxine HCl Riboflavin Thiamine HCl
1. Parker R. C, et al (1957) Special publications, N.Y. Academy of Sciences, 5, 303 a. Values established by the Tissue Culture Association Standards Committee. Tween 80 is a registered trademark of I.CI Americas, Inc *also known as Nicotinamide
Eagle, H (1965) Proc. Soc . Exp. Biol. Med. 89. 362 *also known as Nicotinamide
50
F-12 Nutrient Mixture (Ham) Kaighn's Modification1.2

Base cat. no. Component Inorganic salts CaCl2 (anhyd.) KH4 PO4 CuSO4 5H2O FeSO4 7H2O KCI MgCI2 (anhyd.) MgSO4 NaCI NaHCO3 Na2 HPO4 (anhyd) ZnSO4 7H2O Other components D-Glucose Hypoxanthine Na Lipoic Acid Phenol Red Putrescine 2HCI Sodium Pyruvate Thymidine Amino acids L-Alanine L-Arginine HCI L-Asparagine H2O L-Aspartic Acid L-Cystine HCI H2O L-Glutamine Acid L-Glutamine Glycine L-Histidine HCI H2O L-Isoleucine L-Leucine L-Lysine HCI L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine 2 Na 2H2O L-Valine Vitamins Biotin D-Calcium pantothenate Choline Chloride Folic Acid i-Inositol Niacinamide* Pyridoxine HCI 0.07 0.50 14.00 1.30 18.00 0.037 0.06 18.00 422.00 30.00 26.00 70.00 29.00 292.00 15.00 45.80 7.88 26.20 73.00 8.96 9.92 69.00 21.00 23.00 2.04 13.50 23.00 1260.00 4.00 0.21 3.00 0.32 220.00 0.70 21127 Powder, mg/L 102.00 59.00 0.002 0.80 285.00 49.70 192.00 7530.00 2500.00 115.50 0.144 Riboflavin Thiamine HCI Vitamin B12 0.04 0.30 1.40
1. Ham, RG. (1965) Proc. Nat. Arad.Sci., 53, 288. 2. Kaighn, M. E., Tissue Culture Methods and Applications, (1973), Kruse, P.F., and Patterson Jr M.D. ed p.54-58 *also known as Nicotinamide
Brinster s BMOC-3 Medium
Base cat. no. Component Inorganic salts NaCl KCl CaCl2 KH2PO4 MgSO4 7H2O Glucose Sodium bicarbonate DL-Lactic acid Na Sodium pyruvate Bovine albumin fraction V
11126 1X liquid, mg/L 5546.00 356.00 189.00 162.00 294.00 1000.00 2106.00 2.773 56.00 5000.00
Basal Media 51
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Dulbeccos Modified Eagle Media (DMEM)1

Base cat. no. Component Inorganic salts CaCl2 2H2O CaCl2 (anhyd.) Fe(NO3)3 9H2O KCl MgSO4 (anhyd.) MgSO4 7H2O NaCl NaHCO3 NaH2PO4 2H2O NaH2PO4 H2O* Other components D-Glucose Phenol red HEPES Sodium pyruvate Amino acids L-Alanine L-Aspargine L-Arginine HCl L-Apsrtic Acid L-Cystine L-Cystine 2HCl L-Glutamic Acid L-Glutamine L-Alanyl-L-glutamine Glycyl-L-Glutamine Glycine L-Histidine HCl H2O Amino acids, continued L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine 2Na 2H2O L-Valine Vitamins D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxine HCl Pyridoxal HCI Riboflavin Thiamine HCl 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 105.00 105.00 146.00 66.00 42.00 95.00 16.00 104.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 35.60 60.00 84.00 53.00 48.00 59.00 30.00 42.00 84.00 48.00 30.00 42.00 30.00 42.00 84.00 63.00 84.00 63.00 584.00 30.00 42.00 84.00 63.00 584.00 30.00 42.00 84.00 63.00 584.00 30.00 42.00 84.00 30.00 42.00 30..00 42.00 30..00 42.00 84.00 63.00 584.00 84.00 63.00 4500.00 17.00 1000.00 110.00 4500.00 15.00 15.00 4500.00 15.00 4,500.00 15.00 110.00 4,500.00 15.00 4500.00 5958.00 4500.00 15.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 2644.00 0.10 400.00 97.67 200.00 6400.00 3700.00 141.00 200.00 0.10 400.00 97.67 6400.00 3700.00 125.00 200.00 0.10 400.00 97.67 4750.00 3700.00 125.00 200.00 0.10 400.00 97.67 4750.00 3700.00 200.00 0.10 400.00 97.67 6,400.00 125.00 200.00 0.10 400.00 97.67 6400.00 3700.00 125.00 200.00 0.10 400.00 97.67 4750.00 3700.00 125.00 0.10 400.00 97.67 6400.00 3700.00 125.00 10938 1X liquid, mg/L 11880 1X liquid, mg/L 11960 1X liquid, mg/L 11966 1X liquid, mg/L 11971 1X liquid, mg/L 12800 Powder, mg/L 21013 1X liquid, mg/L 21063 1X liquid, mg/L 21068 1X liquid, mg/L
52
1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396, Smith, J. D,. Freeman, G,. Vogt, M. And Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Association Standards Committee, In Vitro 6:2, 93. a. Values shown are in conformance with the Tissue Culture Association Standards Committee, In Vitro 6:2 93. *also known as Nicotinamide

Base cat. no. Component Inorganic salts CaCl2 2H2O CaCl2 (anhyd.) Fe(NO3)3 9H2O KCl MgSO4 (anhyd.) MgSO4 7H2O NaCl NaHCO3 NaH2PO4 2H2O NaH2PO4 H2O* Other components D-Glucose Phenol red HEPES Sodium pyruvate Amino acids L-Alanine L-Asparagine L-Arginine HCl L-Cystine L-Cystine 2HCl L-Glutamine L-Alanyl-L-glutamine Glycine L-Histidine HCl H2O L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine 2Na 2H2O L-Valine Vitamins D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxine HCI Pyridoxal HCI Riboflavin Thiamine HCl 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 0.40 4.00 4.00 4.00 4.00 7.20 4.00 4.00 w 0.40 4.00 84.00 48.00 862.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 84.00 48.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 84.00 48.00 580.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 84.00 63.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.00 94.00 84.00 62.77 584.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 104.18 94.00 84.00 48.00 580.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 84.00 48.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 84.00 48.00 862.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 84.00 48.00 862.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 94.00 1000.00 15.00 110.00 4500.00 15.00 110.00 1000.00 15.00 5958.00 110.00 4500.00 1000.00 15.00 110.00 1000.00 15.00 110.00 4,500.00 15.00 4500.00 15.00 110.00 4500.00 15.00 5958.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 264.00 0.10 400.00 200.00 3500.00 3700.00 141.00 200.00 0.10 400.00 97.67 6400.00 3700.00 125.00 200.00 0.10 400.00 97.67 6400.00 125.00 264.00 0.10 400.00 200.00 6,400.00 3700.00 141.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 21885 1X liquid, mg/L 21969 1X liquid, mg/L 22320 1X liquid, mg/L 31053 1X liquid, mg/L 31600 1X liquid, mg/L 31885 1X liquid, mg/L 31965 1X liquid, mg/L 31966 1X liquid, mg/L 32430 1X liquid, mg/L
Basal Media 53
continued
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Base cat. no. Component Inorganic salts CaCl2 2H2O CaCl2 (anhyd.) Fe(NO3)3 9H2O KCl MgSO4 (anhyd.) MgSO4 7H2O NaCl NaHCO3 NaH2PO4 2H2OO2 NaH2PO4 H2Oa Other components D-Glucose Phenol red HEPES Sodium pyruvate Amino acids L-Arginine L-Asparagine L-Arginine HCI L-Aspartic acid L-Cystine L-Cystine 2HCl L-Glutamine acid L-Glutamine L-Alanyl-L-Glutamine Glycyl-L-Glutamine Glycine L-Histidine HCl H2O L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine Amino acids, continued L-Tyrosine (disodium salt) L-Valine Vitamins D-Ca pantothenate Choline chloride Folic acid 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 94.00 94.00 94.00 104.20 94.00 94.00 84.00 48.00 580.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 84.00 48.00 580.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 84.00 48.00 580.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 84.00 62.75 580.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 84.00 48.00 580.00 862.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 42.00 95.00 16.00 72.00 4500.00 15.00 4500.00 15.00 110.00 4500.00 15.00 5958.00 4500.00 15.00 4500.00 15.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 264.00 0.10 400.00 200.00 3500.00 3700.00 141.00 200.00 0.10 400.00 97.67 6400.00 125.00 264.00 0.10 400.00 200.00 6400.00 3700.00 141.00 41965 1X liquid, mg/L 41966 1X liquid, mg/L 42430 1X liquid, mg/L 52100 1X Powder, mg/L 61965 1X liquid, mg/L
54

Base cat. no. Vitamins, continued i-Inositol Niacinamide* Pyridoxine HCI Pyridoxal HCI Riboflavin Thiamine HCI 7.20 4.00 4.00 0.40 4.00 7.20 4.00 4.00 0.40 4.00 7.20 4.00 4.00 0.40 4.00 7.20 4.00 4.00 0.40 4.00 7.20 4.00 4.00 0.40 4.00 41965 41966 42430 52100 61965
* Values shown are in conformance with the Tissue Culture Standards Committee, In Vitro 9:6. 1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J.D., Freeman, G., Vogt, M., and Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 6:2, 93.
3 Basal Media 55
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Advanced D-MEM/F-12
Base cat. no. Component Inorganic salts Calcium Chloride (CaCl2) (anhyd.) Cupric SULFATE (CuSO4-5H2O) Ferric Nitrate (Fe(NO3)3 9H2O Ferric sulfate (FeSO4-7H2O) Magnesium Chloride (anhydrous) Magnesium Sulfate(MgSO4) (anhyd.) Potassium Chloride (KCI) Sodium Bicarbonate (NaHCO2) Sodium Chloride (NaCI) Sodium Phosphate dibasic (Na2 HPO4) anhydrous Sodium Phosphate monobasic (NaH2 PO4-HO2) Zinc sulfate (ZnSO4 7H2O) Other components D-Glucose (Dextrose) Ethanolamine Hypoxanthine Na Linoleic Acid Lipoic Acid Phenol Red Putrescine 2HCI Sodium Pyruvate Thymidine Amino acids Glycine L-Alanine L-Arginine hydrochloride L-Asparagine-H2O L-Aspartic Acid L-Cystine hydrochloride-H2O L-Cystine 2HCI L-Glutamic Acid L-Histidine hydrochloride-H2O L-Isoleucine L-Leucine L-Lysine hydrochloride L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine disodium salt dihydrate L-Valine Vitamins Ascorbic Acid phosphate Biotin Choline chloride D-Calcium pantothenate 2.5 0.0035 8.98 2.24 18.75 4.45 147.5 7.5 6.65 17.56 31.29 7.35 31.48 54.47 59.05 91.25 17.24 35.48 17.25 26.25 53.45 9.02 55.79 52.85 3151 1.9 2.39 0.042 0.105 8.1 0.081 110 0.365 116.6 0.0013 0.05 0.417 28.64 48.84 311.8 2438 6995.5 71.02 62.5 0.864 12634 1X liquid, mg/L Folic Acid i-Inositol Niacinemide Pyridoxine hydrochloride Riboflavin Thiamine hydrochloride Vitamin B12 Proteins AlbuMAX II Human Transferrin (Halo) Insulin Recombinant Full Chain Reducing Agents Glutathione, monosodium Trace Elements Ammonium Metavanadate Manganous Chloride Sodium Selenite 0.0003 0.00005 0.0005 1 400 7.5 10 2.65 12.6 2.02 2 0.219 2.17 0.68
1. Dulbecco, R. And Freeman, G., (1959) Plaque formation by the polyoma virus. Virology 8:396 2. Smith, J. D Freeman, G., Vogt, M., et al., The nucleic acid of polyoma virus. Virology 12:185 3. Morton, H.J. (1970) Asurvey of commercially available tissue culture media. In Vitro 6(2):89 4. Ham, R.G., (1965) Clonal growth of mammalian cells in a chemically defined synthetic medium. Proc. Natl. Acad. Sci,. 53:288
56
F-12 Nutrient Mixture (Ham)1

Base cat. no. Component Inorganic salts CaCl2 (anhyd.) CaCl2 2H2O CuSO4 5H2O FeSO4 7H2O KCI MgCI2 (anhyd.) MgCI2 6H2O NaCI NaHCO3 Na2 HPO4 (anhyd) ZnSO4 7H2O Other components D-Glucose Hypoxanthine Hypoxanthine (sodium salt) Linoleic Acid DL-68-Thioctic Acid Phenol Red Putrescine 2HCI Sodium Pyruvate Thymidine Amino acids L-Alanine L-Arginine HCI L-Asparagine H2O L-Asparagine (free base) L-Aspartic Acid L-Cystine HCI L-Glutamic Acid HCI H2O L-Glutamine Acid L-Glutamine L-Alanyl-LGlutamine Glycine L-Histidine HCI H2O L-Isoleucine L-Leucine L-Lysine HCI L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine 8.90 211.00 15.01 13.30 35.12 14.70 146.00 7.50 20.96 3.94 13.10 36.50 4.48 4.96 34.50 10.50 11.90 2.04 8.90 211.00 13.00 13.00 36.00 14.70 146.00 7.50 21.00 4.00 13.00 36.50 4.50 5.00 34.50 10.50 12.00 2.00 5.40 8.90 211.00 13.00 13.00 36.00 14.70 217.00 7.50 21.00 4.00 13.00 36.50 4.50 5.00 34.50 10.50 12.00 2.00 5.40 1802.00 4.77 0.084 0.21 1.20 0.161 110.00 0.73 1802.00 4.00 0.084 0.20 1.20 0.161 110.00 0.70 1802.00 4.00 0.084 0.20 1.20 0.161 110.00 0.70 33.22 0.003 0.834 223.60 57.22 7599.00 142.04 0.863 44.00 0.0024 0.83 223.60 122.00 7599.00 1176.00 142.00 0.86 44.00 0.0024 0.83 223.60 122.00 7599.00 1176.00 142.00 0.86 21700 Powder, mg/L 21765 1X liquid, mg/L 31765 1X liquid, mg/L L-Tyrosine (disodium salt) L-Valine Vitamins Biotin D-Calcium pantothenatec Choline Chloride Folic Acid i-Inositol Niacinamide* Pyridoxine HCI Riboflavin Thiamine HCI Vitamin B12 0.007 0.48 13.96 1.30 18.00 0.037 0.062 0.038 0.34 1.36 0.0073 0.50 14.0 1.30 18.00 0.036 0.06 0.037 0.30 1.40 0.0073 0.50 14.00 1.30 18.00 0.036 0.06 0.037 0.30 1.40 7.81 11.70 11.70 11.70
* Values shown are in conformance with the Tissue Culture Standards Committee, In Vitro 9:6. 1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J.D., Freeman, G., Vogt, M. and Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 6:2, 93.
Basal Media 57
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Fischers Medium for Leukaemic Cells of Mice1
Base cat. no. Component Inorganic salts CaCI2 2H2O KCI MgCI2 6H2O NaCI NaHCO3 NaH2PO4 2H2O Na2HPO4 (anhyd.) Other components D-Glucose Phenol Red Amino acids L-Arginine HCI L-Asparagine L-Cystine L-Glutamine L-Histidine HCI H2O L-Isoleucine L-Leucine L-Lysine HCI L-Methionine L-Phenylalanine L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins Biotin D-Ca pantothenate Choline Chloride Folic Acid i-Inositol Niacinamide* Pyridoxal HCI Riboflavin Thiamine HCI 0.01 0.50 1.50 10.00 1.50 0.50 0.50 0.50 1.00 18.60 10.00 20.00 204.00 60.00 75.00 30.00 50.00 100.00 60.00 15.00 30.00 10.00 60.00 70.00 1000.00 5.00 91.00 400.00 100.00 8000.00 1125.00 77.00 59.00 21475 1X liquid, mg/L
Glasgow MEM 1, 2 (BHK 21)

Base cat. no. Component Inorganic salts CaCI2 (anhyd.) CaCI2 2H2O Fe(NO3)3 9H20 KCI MgSO4 (anhyd.) MGSO4 7H2O NaCI NaHCO3 NaH2PO4 H2O NaH2PO4 2H2O Other components D-Glucose Phenol Red Amino acids L-Arginine HCI L-Cystine L-Cystine 2HCI L-Glutamine L-Histidine HCI H20 L-Isoleucine L-Leucine L-Lysine HCI L-Methionine L-Phenylalanine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine (disodium salt) L-Valine Vitamins D-Ca pantothenate Choline Chloride Folic Acid i-Inositol Niacinamide* Pyridoxal HCI Riboflavin Thiamine HCI 2.00 2.00 2.00 3.60 2.00 2.00 0.20 2.00 2.00 2.00 2.00 3.60 2.00 2.00 0.20 2.00 42.00 24.00 292.00 21.00 52.00 52.00 73.00 15.00 33.00 47.60 8.00 36.20 46.80 42.00 31.28 292.00 21.00 52.40 52.40 73.10 15.00 33.00 47.60 8.00 52.09 46.80 4500.00 15.00 4500.00 15.00 265.00 0.10 400.00 200.00 6400.00 2750.00 141.00 200.00 0.10 400.00 97.67 6400.00 2750.00 124.00 21710 1X liquid, mg/L 22100 Powder, mg/L
* Values shown are in conformance with the Tissue Culture Standards Committee, In Vitro 9:6. 1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J.D., Freeman, G., Vogt, M. and Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 6:2, 93
58
F-10 Nutrient Mixture (Ham)1

Base cat. no. Component Inorganic salts CaCl2 2H2O CuSO4 5H2O FeSO4 7H2O KCI KH2PO4 MgSO4 7H2O NaCI NaHCO3 Na2 HPO4 (anhyd) ZnSO4 7H2O Other components D-Glucose HEPES Hypoxanthine DL-68-Thioctic Acid Phenol Red Sodium Pyruvate Thymidine Amino acids L-Alanine L-Arginine HCI L-Asparagine L-Aspartic Acid L-Cystine L-Glutamic Acid L-Glutamine L-Alanyl-L-Glutamine L-Glycine L-Histidine HCI H2Ob L-Isoleucine L-Leucine L-Lysine HCI L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine 8.92 211.00 12.98 13.30 25.00 14.70 146.00 7.52 23.00 2.60 13.10 29.30 4.48 4.96 11.50 10.50 3.58 0.60 1.81 3.50 9.00 211.00 15.01 13.00 25.00 14.70 146.00 7.50 23.00 2.60 13.00 29.00 4.50 5.00 11.50 10.50 3.60 0.60 1.80 3.50 9.00 211.00 15.01 13.00 25.00 14.70 217.00 7.50 23.00 2.60 13.00 29.00 4.50 5.00 11.50 10.50 3.60 0.60 1.80 3.50 1100.00 5958.00 4.00 0.20 1.20 110.00 0.73 1100.00 4.08 0.20 1.20 110.00 0.73 1100.00 4.00 0.20 1.20 110.00 0.73 22390 1X liquid, mg/L 44.00 0.0025 0.834 285.00 83.00 153.00 6900.00 1200.00 154.50 0.0288 31550 1X liquid, mg/L 44.00 0.0025 0.834 285.00 83.00 153.00 7400.00 1200.00 154.50 0.0288 41550 1X liquid, mg/L 44.00 0.0025 0.834 285.00 83.00 153.00 7400.00 1200.00 154.50 0.0288 Vitamins Biotin D-Calcium pantothenatec Choline Chloride Folic Acid i-Inositol Niacinamide* Pyridoxine HCI Riboflavin Thiamine HCI Vitamin B12 0.024 0.72 0.70 1.32 0.54 0.62 0.21 0.38 1.01 1.36 0.024 0.70c 0.70 1.32 0.50 0.60 0.20 0.40 1.00 1.40 0.024 0.70c 0.70 1.32 0.50 0.60 0.20 0.40 1.00 1.40
3 Basal Media 59
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Insect Cell Culture Media

Base cat. no. 11595 Graces1 unsupplemented 1X liquid, mg/L 750.00 4100.00 2280.00 2780.00 350.00 1013.00 400.00 55.00 700.00 370.00 670.00 60.00 26680.00 225.00 200.00 700.00 350.00 350.00 22.00 600.00 600.00 650.00 11605 Graces 1 supplemented 1X liquid, mg/L 500.00 4100.00 2280.00 2780.00 350.00 1013.00 400.00 55.00 700.00 370.00 3330.00 670.00 60.00 26680.00 3330.00 225.00 200.00 700.00 350.00 350.00 22.00 600.00 600.00 650.00 11300 Graces 1 powder, mg/L 750.00 4100.00a 1070.00 1360.00 1013.00 400.00 55.00 700.00 370.00 670.00 60.00 26680.00 225.00 200.00 700.00 350.00 350.00 28.68 600.00 600.00 650.00 11667 Graces 1 2X liquid, mg/L 1000.00 5600.00 2140.00 2716.00 700.00 2026.00 800.00 110.00 1400.00 740.00 6660.00 1340.00 120.00 53360.00 6660.00 450.00 400.00 1400.00 700.00 700.00 57.36 1200.00 1200.00 1300.00 11405 IPL-41 1X liquid, mg/L 500.00 0.05 0.55 1200.00 918.00 0.02 2850.00 350.00 1160.00 0.04 0.04 4.40 2500.00 29.60 53.60 1000.00 4.80 1650.00 300.00 800.00 1300.00 1300.00 130.35 1500.00 1000.00 200.00 21720 Schneiders 2 1X liquid, mg/L 794.00 1600.00 450.00 3700.00 2100.00 400.00 1321.00d 100.00 2000.00 200.00 100.00 100.00 2000.00 500.00 400.00 400.00 60.00 100.00 800.00 1800.00 250.00
Component Inorganic salts CaCl2 CaCl2 2H2O CoCl26H2O FeSO47H2O KCl KH2PO4 MgCl2 MgCl2 6H2O MgSO4 MgSO4 7H2O MnCl2 4H2O NaCl NaHCO3 NaH2PO4 H2O NaH2PO4 2H2O NaH2PO4 7H2O (NH4)6 (Mo7 O24 . 4H2O) ZnCl2 Other components Fructose Fumaric acid D-Glucose -Ketoglutaric acid Lactalbumin Hydrolysate Malic acid Maltose Succinic acid Sucrose Trehalose Tryptose Broth Yeastolate Amino acids L-Alanine -Alanine L-Arginine L-Arginine HCl L-Asparagine L-Aspartic acid L-Cysteine L-Cystine L-Cystine 2HCl L-Cystine 2Na L-Glutamic acid L-Glutamine Glycine
1. Grace, T. CC (1962) Nature 195, 788. a. Increased from 2240.00 mg/L upon recommendation of Doctor Vaughan 2. Schneider, Imogene (1964) Exp, Zool, 156:1:9 a. Revision of original formulation, Scneider, Imogen (1965) personal communications b. Original formulations list Na2HPO4 at 700.00 mg/L. Revise formulation believed by author to possess better buffering capacity and better nutritive properties.
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Insect Cell Culture Media

Base cat. no. 11595 Graces1 unsupplemented 1X liquid, mg/L 2500.00 50.00 75.00 625.00 150.00 350.00 1100.00 175.00 100.00 50.00 100.00 0.02 0.01 0.02 0.20 0.02 0.02 0.02 0.02 0.02 0.02 11605 Graces 1 supplemented 1X liquid, mg/L 2500.00 50.00 75.00 625.00 50.00 150.00 350.00 550.00 175.00 100.00 50.00 100.00 0.02 0.01 0.02 0.20 0.02 0.02 0.02 0.02 0.02 0.02 11300 Graces 1 powder, mg/L 2500.00 50.00 75.00 625.00 50.00 150.00 350.00 550.00 175.00 100.00 62.00 100.00 0.02 0.01 0.02 0.20 0.02 0.02 0.02 0.02 0.02 0.02 11667 Graces 1 2X liquid, mg/L 5000.00 100.00 150.00 1250.00 100.00 300.00 700.00 1100.00 350.00 200.00 124.00 200.00 0.04 0.02 0.04 0.40 0.04 0.04 0.04 0.04 0.04 0.04 11405 IPL-41 1X liquid, mg/L 200.00 800.00 750.00 250.00 700.00 1,000.00 1,000.00 500.00 200.00 200.00 100.00 500.00 0.32 0.16 0.008 20.00 0.08 0.40 0.16 0.40 0.08 0.08 0.24 21720 Schneiders 2 1X liquid, mg/L 400.00 150.00 150.00 1650.00 800.00 150.00 1700.00 250.00 350.00 100.00 500.00 300.00
Component Amino acids, continued L-Histidine L-Hydroxyproline L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine 2Na L-Valine Vitamins Para-aminobenzoic acid D-Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacin Pyridoxine HCl Riboflavin Thiamine HCl Vitamin B12
3 Basal Media
1. Grace, T. CC (1962) Nature 195, 788. a. Increased from 2240.00 mg/L upon recommendation of Doctor Vaughan 2. Schneider, Imogene (1964) Exp, Zool, 156:1:9 a. Revision of original formulation, Scneider, Imogen (1965) personal communications b. Original formulations list Na2HPO4 at 700.00 mg/L. Revise formulation believed by author to possess better buffering capacity and better nutritive properties.
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Iscoves Modified Dulbeccos Media (IMDM)1,2,3

Base cat. no. Component Inorganic salts CaCl2 (anhyd.) CaCl2 2H2O KCl KNO3 MgSO4 2H2O MgSO4 (anhyd.) NaCl NaHCO3 NaH2PO4 H2Oc NaH2PO4 2H2O Na2SeO3 5H2O Other components D-Glucose Phenol red HEPES Sodium pyruvate Amino acids L-Alanine L-Asparagine H2O L- Asparagine (freebase) L- Arginine H2O L-Aspartic acid L-Cystine 2HCl L-Cystine L-Glutamic acid L-Glutamine L-Alanyl-L-glutamine Glycine L-Histidine HCl H2O L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine (disodium salt) L-Valine Vitamins Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxal HCl 0.013 4.00 4.00 4.00 7.20 4.00 4.00 0.013 4.00 4.00 4.00 7.20 4.00 4.00 0.013 4.00 4.00 4.00 7.20 4.00 4.00 0.013 4.00 4.00 4.00 7.20 4.00 4.00 25.00 28.40 84.00 30.00 70.00 75.00 584.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 40.00 42.00 95.00 16.00 104.20 94.00 25.00 28.40 84.00 30.00 70.00 75.00 584.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 40.00 42.00 95.00 16.00 104.20 94.00 25.00 28.40 84.00 30.00 70.00 75.00 868.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 40.00 42.00 95.00 16.00 104.20 94.00 25.00 25.00 84.00 30.00 91.24 75.00 584.00 30.00 42.00 105.00 105.00 146.00 30.00 66.00 40.00 42.00 95.00 16.00 104.20 94.00 4500.00 5958.00 110.00 4500.00 15.00 5958.00 110.00 4500.00 15.00 5958.00 110.00 4500.00 15.00 5958.00 110.00 219.00 330.00 0.076 200.00 4505.00 3024.00 141.00 0.017 219.00 330.00 0.076 200.00 4505.00 3024.00 141.00 0.017 219.00 330.00 0.076 200.00 4505.00 3024.00 141.00 0.017 165.00 330.00 0.076 97.67 4500.00 125.00 0.017 20156 1X liquid, mg/L 21980 1X liquid, mg/L 31980 1X liquid, mg/L 42200 Powder, mg/L Riboflavin Thiamine HCl Vitamin B12 0.40 4.00 0.013 0.40 4.00 0.013 0.40 4.00 0.013 0.40 4.00 0.013
1. Dubecco, R. And Freeman, G. (1959) Virology 8, 396. Smith, J. D., Freeman, G., Vogt, M. And Dubecco, R. (1960) Virology 12, 165. Tissue Culture Standards Committee, In Vitro 62 93. 2. Iscove, N, N. Melchers, F,. J. Experimental medicine 147, 923. 3. Iscove, N, N Personal Communication. b. Values shown are in conformance with the tissue culture standards committee, In Vitro (1970) 9.6 * Also known as Nicotindamide.
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Improved MEM Zinc Option (Richters Modification)1

Base cat. no. Component Inorganic salts CaCl2 FeCl3 6H2O KCl MgCl2 MgSO4 NaCl NaHCO3 NaH2PO4 H2O ZnSO4 H2O Other components D-Glucose Gentamicin sulfate Putrescine HCl Sodium pyruvate Lipoic acid Linoleic acid Amino acids L-Arginine H2O L-Asparagine H2O L-Cystine L-Glutamine L-Histidine HCl H2O L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxal HCl Riboflavin Thiamine HCl Vitamin B12
1.Richter, a., Sanford, K., and Evans, V., (1972), J. National Cancer inst. 49, 1705.
Leibovitzs L-15 Media 1

Base cat. no. 11415 1X liquid, mg/L 185.00 400.00 60.00 200.00 8000.00 190.00 900.00 10.00 550.00 225.00 500.00 250.00 120.00 300.00 200.00 250.00 250.00 125.00 75.00 75.00 125.00 200.00 300.00 20.00 300.00 100.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 21083 1X liquid, mg/L 185.00 400.00 60.00 200.00 8000.00 190.00 900.00 550.00 225.00 500.00 250.00 120.00 300.00 200.00 250.00 250.00 125.00 75.00 75.00 125.00 200.00 300.00 20.00 300.00 100.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 31415 1X liquid, mg/L 185.00 400.00 60.00 200.00 8000.00 190.00 900.00 10.00 550.00 225.00 500.00 250.00 120.00 200.00 250.00 250.00 125.00 75.00 75.00 125.00 200.00 300.00 20.00 300.00 100.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 41300 Powder, mg/L 140.00 400.00 60.00 93.68 97.67 8000.00 190.12 900.00 10.00 550.00 225.00 500.00 250.00 120.00 300.00 200.00 250.00 250.00 125.00 75.00 75.00 125.00 200.00 300.00 20.00 300.00 100.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00
10373 1X liquid, mg/L 200.00 0.54 400.00 85.75 12.22 6800.00 2200.00 150.00 0.10 2000.00 0.05 0.20 110.00 0.20 0.10 126.00 60.00 24.00 584.00 42.00 52.00 131.00 73.00 15.00 32.00 2.00 42.00 48.00 10.00 36.00 46.00 0.10 1.00 56.00 1.00 36.00 1.00 1.00 0.10 1.00 0.20 Component Inorganic salts CaCl2 (anhyd.) CaCl2 2H2O KCl KH2PO4 MgCl2 (anhyd.) MgCl2 6H2O MgSO4 (anhyd.) NaCl Na2HPO4 Other components D(+)-Galactose Phenol red Sodium pyruvate Amino acids L-Alanine L-Arginine (free base) L-Asparagine L-Cysteine (free base) L-Glutamine Glycine L-Histidine (free base) L-Isoleucine L-Leucine L-Lysine (free base) L-Methionine L-Phenylalanine L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxal HCl Pyridoxine HCl Flavin Monoucleotide Thiamine Monophosphate HCI
3 Basal Media 63
b. Mono hydrate Form listed by the Tissue Culture Standards Committee, In Vitro (1974) 9..6. * Also known as Nicotindamide.
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McCoys 5A Media (modified) 1 ,2,3

Base cat. no. Component Inorganic salts CaCl2 2H2O KCl MgSO4 7H2O NaCl NaHCO3 NaH2PO4 2H2O Other components Bacto-peptone HEPES D-Glucose Glutathione (reduced) Phenol red Amino acids L-Alanine L-Arginine HCl L-Asparagine L-Cysteineb L-Glutamic Acid L-Glutamine L-Alanyl-L-Glutamine Glycine L-Histidine HCl H2O L-Hydroxyproline L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins Ascorbic acid Biotin Choline chloride D-Ca pantothenate Folic acid i-Inositol Niacin Niacinamide Para-aminobenzoic acid Pyridoxine HCl Riboflavin Thiamine HCl Vitamin B12 0.50 0.20 5.00 0.20 9.80 36.00 0.50 0.50 1.00 0.50 0.20 0.20 2.3075 0.50 0.20 10.00 0.20 10.00 36.00 0.50 0.50 1.00 0.50 0.20 0.20 2.00 0.50 0.20 10.00 0.20 10.00 36.00 0.50 0.50 1.00 0.50 0.20 0.20 2.00
a
MCDB 1 31 Medium
36600 1X liquid, mg/L 132.00 400.00 200.00 6460.00 2200.00 649.00 600.00 3000.00 0.50 10.00 13.40 42.10 45.00 19.90 31.50 22.10 326.00 7.50 20.90 19.70 39.30 39.30 36.50 14.90 16.50 17.30 26.30 17.90 3.10 18.10 17.60 Base cat. no. Component Inorganic salts CuSO4 5H2O MnSO4 H2O ZnSO4 H2O NiCl2 6H2O FeSO4 7H2O Na2SiO3 9H2O CaCl2 2H2O KCl MgSO4 7H2O NaCl NaHCO3 Na2HPO4 7H2O H2SeO3 (NH4)6Mo7O24 4H2O NH4VO3 Other components D-Glucose Adenine Thymidine Phenol red Sodium pyruvate Lipoic acid Putrescine 2HCl Amino acids L-Alanine L-Arginine HCl L-Asparagine H2O L-Aspartic acid L-Cysteine 2HCl H2O L-Glutamic acid Glycine L-Histidine HClH2O L-Isoleucine L-Leucine L-LysineHCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins Biotin D-Ca pantothenate Choline Chloride Folinic Acid i-Inositol Niacinamide* Pyridoxine HCl Riboflavin Thiamine HCl Vitamin B12
* Also known as Nicotindamide
22330 1X liquid, mg/L 132.00 400.00 200.00 5960.00 2200.00 655.00 600.00 5960.00 3000.00 0.50 10.00 13.40 42.10 45.00 19.90 31.50 22.10 219.20 7.50 20.90 19.70 39.30 39.30 36.50 14.90 16.50 17.30 26.30 17.90 3.10 18.10 17.60
26600 1X liquid, mg/L 132.00 400.00 200.00 6460.00 2200.00 649.00 600.00 3000.00 0.50 10.00 13.40 42.10 45.00 19.90 31.50 22.10 219.20 7.50 20.90 19.70 39.30 39.30 36.50 14.90 16.50 17.30 26.30 17.90 3.10 18.10 17.60
10372 1X liquid, mg/L 0.0012 0.0002 0.0003 0.0007 0.283 2.80 235.00 298.00 2464.00 6430.00 1176.00 134.00 0.0038 0.0037 0.0006 1000.00 0.135 0.024 12.40 110.00 0.0021 0.002 2.70 63.20 15.00 13.30 35.00 4.00 2.30 42.00 66.00 131.00 182.00 15.00 33.00 11.50 32.00 12.00 4.10 18.10 12.00 0.0073 12.00 1.40 0.60 7.20 6.10 2.10 0.0038 3.40 0.0036
L-Aspartic acid
1. McCoy, T. A., Maxwell, M. And Kruse, P, F. (1995) Proc. Soc. Exper, Biol, Med., 100. 115. 2. Hsu. T. C. And kellog, D, S, Jr (1960) J. Nat. Cancer inst., 25.221 3. Iwakata, S. And Grace, J , T., Jr. (1964) N. Y. J. Med., 64:18, 2279 a. HCI Form listed by the Tissue Culture Standards Committee, In Vitro (1974) 9..6. b. Mono hydrate Form listed by the Tissue Culture Standards Committee, In Vitro (1974) 9:6. * Also known as Nicotindamide.
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Media 199 1 with Earle's Salts

Base cat. no. Component Inorganic salts CaCl2 H2O CaCl2 (anhyd) Fe(NO3)3 9H2O KCl KH2PO4 MgSO4 7H2O MgSO4 (anhyd.) NaCl NaHCO3 Na2HPO4 2H2O NaH2PO4 H2O Other components Adenine sulfate Adenosine-5-triphosphate Adenosine-5-phosphate Cholesterol Deoxyribose D-Glucose Glutathione (reduced) Guanine Guanine HCl HEPES Hypoxanthine Hypoxanthine Na Phenol red Ribose Sodium Acetate 3H2O Sodium Acetate Thymine Tween 80 Uracil Xanthine Xanthine Na Amino acids L-Alanine L-Arginine HCl L-Aspartic Acid L-Cysteine HCl H2O L-Cystine HCl L-Cystine L-Cystine 2HCl L-Glutamic acid 25.00 70.00 30.00 0.10 26.00 75.00 50.00 140.00 60.00 22.00 40.00 119.07 250.00 700.00 300.00 1.10 200.00 592.43 25.00 70.00 30.00 0.11 20.00 59.54 25.00 70.00 30.00 0.11 26.06 66.82 25.00 70.00 30.00 0.10 20.00 59.24 25.00 70.00 30.00 0.10 20.00 59.24 25.00 70.00 30.00 0.10 20.00 59.24 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 0.40 0.50 50.0 0.30 20.00 0.30 0.34 20.00 2.00 0.40 0.40 1.00 2000.00 0.10 0.60 0.60 1.00 166.00 0.60 40.00 0.60 0.60 100.00 10.00 2.00 2.00 5.00 10000.00 0.50 3.00 3.00 200.00 5.00 830.00 3.00 200.00 3.00 3.00 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 5960.00 0.30 20.00 0.50 83.0 0.30 20.00 0.30 0.30 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 0.40 0.354 20.00 0.50 50.00 0.30 20.00 0.30 0.344 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 20.00 0.50 83.00 0.30 20.00 0.30 0.30 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 0.30 20.00 0.50 83.00 0.30 20.00 0.30 0.30 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 0.30 20.00 0.50 83.00 0.30 20.00 0.30 0.30 200.00 0.72 400.00 400.00 98.00 6800.00 2200.00 140.00 528.00 1.40 800.00 13600.00 2500.00 316.00 2640.00 7.20 4000.00 2000.00 6800.00 1,580.00 264.00 0.10 400.00 200.00 6300.00 2200.00 158.00 200.00 0.72 400.00 97.67 6.800.00 140.00 264.00 0.70 400.00 200.00 6800.00 2200.00 158.00 264.00 0.70 400.00 200.00 6800.00 1250.00 140.00 264.00 0.70 400.00 200.00 6800.00 2200.00 158.00 11043 1X liquid, mg/L 21157 2X liquid, mg/L 21180 10X liquid, mg/L 22340 1X liquid, mg/L 31100 Powder, mg/L 31150 1X liquid, mg/L 31153 1X liquid, mg/L 41150 1X liquid, mg/L
3 Basal Media
1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J. D., Freeman, G., Vogt, M. And Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 62, 93. a. Value shown are in conformance with the Tissue Culture Standards Committee, In Vitro 96 *Also known as Nicotindamide.
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Media 199 1 with Earle's Salts

Base cat. no. Component Amino acids, continued L-Glutamine Glycine L-Histidine HCl H2O L-Hydroxyproline L-Isoleucine L-Leucine L-LysineHCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine 2Na 2H2O L-Valine Vitamins Ascorbic acid -Tocopherol phosphate (sodium salt) Biotin Calciferol D-Ca pantothenate Choline chloride Folic acid i-Inositol Menadione Niacin Niacinamide* Para-aminobenzoic Acid Pyridoxal HCl Pyridoxine HCl Riboflavin Thiamine HCl Vitamin A (acetate)a 0.05 0.01 0.01 0.10 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.14 0.10 0.02 0.20 0.02 1.00 0.02 0.10 0.02 0.05 0.025 0.10 0.05 0.05 0.02 0.02 0.20 0.50 0.10 0.10 1.00 0.10 5.00 0.10 0.50 0.10 0.25 0.30 0.50 0.30 0.30 0.10 0.10 1.00 0.05 0.01 0.01 0.10 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.10 0.05 0.01 0.01 0.10 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.14 0.05 0.01 0.01 0.10 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.10 0.05 0.01 0.01 0.10 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.10 0.05 0.01 0.01 0.10 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.10 100.00 50.00 22.00 10.00 40.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 58.00 25.00 200.00 100.00 43.76 20.00 80.00 120.00 140.00 30.00 50.00 80.00 50.00 60.00 20.00 80.00 50.00 1000.00 500.00 218.80 100.00 400.00 600.00 700.00 150.00 250.00 400.00 250.00 300.00 100.00 400.00 250.00 100.00 50.00 21.88 10.00 40.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 40.00 25.00 100.00 50.00 21.88 10.00 40.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 57.88 25.00 100.00 50.00 21.88 10.00 40.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 40.00 25.00 100.00 50.00 21.88 10.00 40.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 40.00 25.00 50.00 21.88 10.00 40.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 40.00 25.00 11043 1X liquid, mg/L 11043 2X liquid, mg/L 11043 10X liquid, mg/L 11043 1X liquid, mg/L 11043 Powder, mg/L 11043 1X liquid, mg/L 31153 1X liquid, mg/L 41150 1X liquid, mg/L
1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J. D., Freeman, G., Vogt, M. And Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 62, 93. a. Value shown are in conformance with the Tissue Culture Standards Committee, In Vitro 96 * Also known as Nicotindamide.
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Minimum Essential Medium (MEM) Media 1

Base cat. no. Component Inorganic salts CaCl2 (anhyd.) CaCl2 2H2O KCl MgSO4 (anhyd.) MgSO4 7H2O NaCl NaHCO3 Na2HPO4 H2O Na2HPO4 2H2O Other components D-Glucose DL-68 Thioctic Acid Phenol red Sodium Pyruvate Amino acids L-Alanine L-Arginine HCl L-Asparagine H2O L-Aspartic acid L-Cystine L-Cystine 2HCl L-Cysteine HCl L-Cysteine HCl H2O L-Glutamic acid L-Glutamine L-Alanyl-L-glutamine Glycine L-Histidine HCl H2O L-Isoleucine L-Leucine L-LysineHCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine (disodium salt) L-Valine 25.00 126.98 50.00 30.00 31.00 100.00 75.00 292.00 50.00 41.88 52.40 52.40 72.47 15.00 32.00 40.00 25.00 48.00 10.00 52.09 46.00 25.00 127.00 25.00 30.00 24.00 100.00 75.00 292.00 50.00 42.00 53.00 52.00 73.00 15.00 32.00 40.00 25.00 48.00 10.00 36.00 46.00 25.00 127.00 50.00 30.00 24.00 100.00 75.00 292.00 50.00 42.00 53.00 52.00 73.00 15.00 32.00 40.00 25.00 48.00 10.00 36.00 46.00 25.00 127.00 25.00 30.00 24.00 100.00 75.00 434.00 50.00 42.00 53.00 52.00 73.00 15.00 32.00 40.00 25.00 48.00 10.00 36.00 46.00 25.00 127.00 50.00 30.00 24.00 100.00 75.00 434.00 50.00 42.00 53.00 52.00 73.00 15.00 32.00 40.00 25.00 48.00 10.00 36.00 46.00 25.00 127.00 50.00 30.00 31.00 100.00 75.00 292.00 50.00 42.00 53.00 52.00 73.00 15.00 32.00 40.00 25.00 48.00 10.00 36.00 46.00 1000.00 0.20 10.00 110.00 1000.00 0.20 10.00 110.00 1000.00 0.20 10.00 110.00 1000.00 0.20 10.00 110.00 1000.00 0.20 10.00 110.00 1000.00 0.20 110.00 200.00 400.00 97.67 6800.00 140.00 264.00 400.00 200.00 6800.00 2200.00 158.00 264.00 400.00 200.00 6800.00 2200.00 158.00 264.00 400.00 200.00 6800.00 2200.00 158.00 264.00 400.00 200.00 6800.00 2200.00 158.00 264.00 400.00 200.00 6800.00 2200.00 158.00 11900 Powder, mg/L 22561 1X liquid, mg/L 22571 1X liquid, mg/L 32561 1X liquid, mg/L 32571 1X liquid, mg/L 41061 1X liquid, mg/L
3 Basal Media
1. Nature, new biology (1971) 230, 52.
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Base cat. no. Component Vitamins L-Ascorbic acid Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxal HCl Riboflavin Thiamine HCl Vitamin B12 Ribonucleosides Adenosine Cytidine Guanosine Uridine Deoxyribonucleosides 2 Deoxyadenosine 2 Deoxycytidine HCl 2 Deoxyguanosine 2 Deoxythymidine
11900 Powder, mg/L 50.00 0.10 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.36 10.00 10.00 10.00 10.00 10.00 11.00 10.00 10.00
22561 1X liquid, mg/L 50.00 0.10 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.40
22571 1X liquid, mg/L 50.00 0.10 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.40 10.00 10.00 10.00 10.00 10.00 11.00 10.00 10.00
32561 1X liquid, mg/L 50.00 0.10 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.40
32571 1X liquid, mg/L 50.00 0.10 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.40 10.00 10.00 10.00 10.00 10.00 11.00 10.00 10.00
41061 1X liquid, mg/L 50.00 0.10 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.40 10.00 10.00 10.00 10.00 10.00 11.00 10.00 10.00
1. Nature, new biology (1971) 230, 52.
68
Medium 1991 with Hanks' Salts

Base cat. no. Component Inorganic salts CaCl2 H2O CaCl2 (anhyd.) Fe(NO3)3 9H2O KCl KH2PO4 MgSO4 7H2O MgSO4 (anhyd.) NaCl NaHCO3 Na2HPO4 2H2O Na2HPO4 H2O Na2HPO4 (anhyd.) Other components Adenine Sulphate Adenosine-5-triphosphate Adenosine-5-phosphate Cholesterol Dexoxyriboxe D-Glucose Glutathione (reduced) Guanine Guanine HCI HEPES Hypoxanthine Hypoxanthine Na Phenol Red Ribose Sodium Acetate 3H2O Sodium Acetate Thymine Tween 80 Uracil Xanthine XanthineNa Amino acids L-Alanine L-Arginine HCl L-Aspartic acid L-Cysteine HCl H2O L-Cysteine HCl L-Cystine L-Cystine 2HCl L-Glutamic acid L-Glutamine L-Alanyl-L-glutamine Glycine L-Histidine HCl H2O L-Hydroxyproline 25.00 70.00 30.00 0.11 26.06 66.82 100.00 50.00 21.88 10.00 25.00 70.00 30.00 0.11 20.00 59.54 100.00 50.00 21.88 10.00 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 5958.00 0.344 20.0 50.0 50.00 30.0 20.0 30.0 0.344 10.00 1.00 0.20 0.20 0.50 1000.00 0.05 0.30 5960.00 0.30 20.0 50.0 83.00 30.0 200.00 30.0 30.0 140.10 0.72 400.00 60.00 97.70 7500.00 47.40 185.00 0.10 400.00 60.00 200.00 7500.00 350.00 48.00 10012 Powder, mg/L 22350 1X Liquid, mg/L L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Thyptophan L-Thyrosine L-Thyrosine Na 2H2O L-Valine Vitamins Ascorbic Acid - Tocopherol Phosphate (disodium salt) Biotin Calciferol D-Ca Pantothenate Choline Chloride Folic Acid i-Inositol Menadione Niacin Niacinamide* Para-aminobenzoic Acid Pyridoxal HCI Pyridoxine HCI Riboflavin Thiamine HCI Vitamin A (acetate)a 0.05 0.01 0.01 10.00 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.14 0.05 0.01 20.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 57.88 25.00 20.00 60.00 70.00 15.00 25.00 40.00 25.00 30.00 10.00 40.00 25.00
3 Basal Media
0.01 10.0 0.01 0.50 0.01 0.05 0.01 0.025 0.025 0.05 0.025 0.025 0.01 0.01 0.10
1. Dulbecco, R., Freeman, G. (1959) Virology 8, 396. Smith, J. D., Freeman, G., Vogt, M. And Dulbecco, R. (1960) Virology 12, 185. Tissue Culture Standards Committee, In Vitro 62, 93. a. Value shown are in conformance with the Tissue Culture Standards Committee, In Vitro 96 * Also known as Nicotindamide.
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Modified Eagle Medium (MEM), 2X 1 with Earles Modified Salts

Base cat. no. Component Inorganic salts CaCl2 2H2O Fe(NO3)3 9H2O KCl MgSO47H2O NaCl NaHCO3 NaH2PO4 H2O Other components D-Glucose Amino acids L-Arginine L-Cystine L-Glutamine L-Histidine L-Isoleucine L-Leucine L-Lysine L-Methionine L-Phenylalanine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide Riboflavin Thiamine HCl
*
NCTC-109 Medium
Base cat. no. Component Inorganic salts CaCl2 200.00 400.00 97.67 6800.00 2200.00 140.00 1000.00 1.80 20.00 50.00 1.80 12.50 2.50 7.00 1.00 1.00 1.00 1.00 50.00 10.00 10.00 10.00 10.00 0.10 10.00 31.60 5.50 31.20 9.20 9.90 14.00 290.00 3.90 KCl MgSO4 NaCl NaHCO3 NaH2PO4 H2O Other components D-Glucose D-Glucuronolactone Phenol red (sodium salt) Sodium acetate Sodium glucuronate H2O Tween 80 Coenzymes CoA (coenzyme A) DPN (diphosphopyridine nucleotide) FAD (flavin adenine dinucleotide) TPN (triphosphopyridine nucleotide monosodium) TPP (cocarboxylase) UTP (uridine triphosphate, sodium) Reducing agents Ascorbic acid Glutathione, monosodium Nucleic acid derivatives Deoxyadenosine Deoxyguanosine Deoxycytidine HCl 5-Methylcytosine Thymidine Amino acids L-Alanine L--Amino-n-butyric acid L-Arginine HCl L-AsparagineH2O L-Aspartic acid L-Cystine2HCl L-CystineHCIH2O D-GlucosamineHCl 21340 1X liquid, mg/L Base cat. no. Component Amino acids, continued L-Glutamic acid Glycine L-Histidine HCl H2O Hydroxy-L-Proline L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Ornithine HCl L-Phenylalanine L-Proline L-Serine L-Taurine L-Threonine L-Tryptophan L-Tyrosine 2Na 2H2O L-Valine Vitamins -Tocopherol phosphate (disodium salt) d-Biotin Calciferol (Vitamin D2) D-Ca pantothenate Choline chloride Folic acid i-Inositol Menadione (Vitamin K) Niacin Niacinamide* Para-aminobenzoic acid Pyridoxal HCl Pyridoxine HCl Riboflavin Thiamine HCl Vitamin A (crystallized alcohol) Vitamin B12
* Also known as Nicotindamide. Tween 80 is a registered trademark of ICI Americas
21340 1X liquid, mg/L 8.30 13.60 26.70 4.10 18.00 20.40 38.60 4.50 9.40 16.50 6.20 10.80 4.20 18.90 17.50 21.00 25.00 0.025 0.025 0.250 0.025 1.25 0.025 0.125 0.025 0.063 0.063 0.125 0.063 0.063 0.025 0.025 0.25 10.00
21935 2X liquid, mg/L 376.00 1.00 800.00 400.00 12800.00 8400.00 238.00 9000.00 420.00 96.00 870.00 124.00 210.00 209.60 232.00 60.00 128.00 192.00 40.00 144.00 184.00 4.00 4.00 4.00 8.00 4.00 4.00 0.40 4.00
Pyridoxal HCl
1. Daniel, M. D. And Melendez V. V. (1969) Proc, Soc, Exp, Biol, Med. 127:33. * Also known as Nicotindamide.
70
Minimum Essential Media1 (MEM)

Base cat. no. Component Inorganic salts CaCl2 2H2O CaCl2 (anhyd.) KCl KH2PO4 MgSO4 (anhyd.) MgSO4 7H2O MgCI2 6H2O NaCl NaHCO3 NaH2PO4 2H2O NaH2PO4 H2Oa NaH2PO4 Other components D-Glucose Phenol red Sodium succinate Succinic acid HEPES Amino acids L-Alanine L-Arginine HCl L-Asparagine H2O L-Aspartic acid L-Cystine L-Cystine2HCl L-Glutamic acid L-Glutamine Glycine L-Histidine HCl H2O L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine (disodium salt) L-Valine Vitamins D-Ca pantothenate Choline bitartrate Choline chloride Folic acid i-Inositol Niacinamide* 1.00 1.00 1.00 2.00 1.00 1.00 1.00 1.00 2.00 1.00 1.00 1.00 1.00 2.00 1.00 1.00 1.80 1.00 2.00 1.00 1.00 1.00 1.00 2.00 1.00 1.00 1.00 1.00 2.00 1.00 10.00 10.00 10.00 20.00 10.00 1.00 1.00 1.00 2.00 1.00 9.00 126.00 13.00 13.00 31.00 15.00 7.50 42.00 52.00 52.00 72.50 15.00 32.00 12.00 11.00 48.00 10.00 52.00 46.00 126.00 31.29 292.00 42.00 52.00 52.00 72.50 15.00 32.00 48.00 10.00 52.09 46.00 126.00 31.00 42.00 52.00 52.00 73.00 15.00 32.00 48.00 10.00 52.00 46.00 126.00 31.00 42.00 52.00 52.00 72.50 15.00 32.00 48.00 10.00 36.00 52.09 46.00 126.00 31.29 292.00 42.00 52.00 52.00 72.50 15.00 32.00 48.00 10.00 52.00 46.00 126.00 24.00 42.00 52.00 52.00 73.00 15.00 32.00 48.00 10.00 36.00 46.00 126.00 24.00 42.00 52.00 52.00 73.00 15.00 320.00 480.00 100.00 360.00 46.00 126.00 24.00 292.00 42.00 52.00 52.00 73.00 15.00 32.00 48.00 10.00 36.00 46.00 1000.00 10.00 1000.00 10.00 5958.00 1000.00 10.00 1000.00 6.00 100.00 75.00 1000.00 10.00 5958.00 1000.00 10.00 1000.00 10.00 1000.00 10.00 200.00 400.00 98.00 6800.00 2200.00 140.00 140.00 400.00 60.00 97.67 7500.00 47.88 400.00 98.00 6800.00 2200.00 1400.00 200.00 400.00 97.67 6800.00 140.00 200.00 400.00 97.67 6300.00 140.00 264.00 200.00 400.00 200.00 6800.00 2200.00 158.00 2640.00 400.00 2,000.00 6,800.00 1580.00 185.00 400.00 60.00 100.00 100.00 8000.00 350.00 48.00 10370 1X liquid, mg/L 11012 Powder, mg/L 11380 1X liquid, mg/L 11700 Powder, mg/L 21011 Powder, mg/L 21090 1X liquid, mg/L 21430 10X liquid, mg/L 21575 1X liquid, mg/L
3 Basal Media
1.Eagle, H. (1959) Science, 130, 432. a. Original Formula lists this compnent as NaH2PO4, 2H2O2 * Also known as Nicotindamide.
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Vitamins, continued Pyridoxal HCl Riboflavin Thiamine HCl 1.00 0.10 1.00 1.00 0.10 1.00 1.00 0.10 1.00 1.00 0.10 1.00 1.00 0.10 1.00 1.00 0.10 1.00 10.00 1.00 10.00 1.00 0.10 1.00
1.Eagle, H. (1959) Science, 130, 432. a. Original Formula lists this compnent as NaH2PO4, 2H2O2 * Also known as Nicotindamide.
72
Minimum Essential Media1 (MEM)

Base cat. no. Component Inorganic salts CaCl22H2O CaCl2 (anhyd.) KCl MgSO4 (anhyd.) MgSO47H2O MgCI26H2O NaCl NaHCO3 NaH2PO42H2O NaH2HPO42H2Oa Other components D-Glucose HEPES Phenol red Amino acids L-Alanine L-Arginine HCl L-Asparagine H2O L-Aspartic acid L-Cystine L-Cystine2HCl L-Glutamic acid L-Glutamine L-Alanyl-L-glutamine Glycine L-Histidine HClH2O L-Isoleucine L-Leucine L-LysineHCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine (disodium salt) L-Valine Vitamins D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Pyridoxal HCl Riboflavin Thiamine HCl 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 1.00 1.00 1.00 2.00 1.00 1.00 0.10 1.00 126.40 24.02 292.00 41.92 52.46 52.46 73.06 14.92 33.02 47.64 10.20 36.22 46.86 126.40 24.02 41.92 52.46 52.46 73.06 14.92 33.02 47.64 10.20 36.22 46.86 126.00 24.02 406.00 41.92 52.46 52.46 73.06 14.92 32.02 47.64 10.20 36.22 46.86 8.90 126.40 13.02 13.30 31.29 14.70 292.00 7.50 42.00 52.00 52.00 72.50 15.00 32.00 11.50 10.50 48.00 10.00 52.09 46.00 126.40 24.02 434.00 41.92 52.46 52.46 73.06 14.92 33.02 47.64 10.20 36.22 46.86 126.40 24.02 41.92 52.46 52.46 73.06 14.92 33.02 47.64 10.20 36.22 46.86 126.00 31.29 292.00 42.00 52.00 52.00 72.50 15.00 32.00 48.00 10.00 51.09 46.00 1,000.00 10.00 1,000.00 5,960.00 10.00 1,000.00 10.00 1,000.00 10.00 1,000.00 5,960.00 10.00 1,000.00 1,000.00 10.00 200.00 6,800.00 2,200.00 158.00 264.00 400.00 264.00 400.00 200.00 6,300.00 2,200.00 158.00 200.00 6,800.00 2,200.00 158.00 264.00 400.00 264.00 400.00 97.67 6,800.00 140.00 200.00 6,300.00 2,200.00 158.00 264.00 400.00 264.00 400.00 200.00 6,800.00 2,200.00 158.00 200.00 400.00 97.67 6,800.00 140.00 31095 1X liquid, mg/L 32360 1X liquid, mg/L 41090 1X liquid, mg/L 41500 Powder, mg/L 42360 1X liquid, mg/L 51200 1X liquid, mg/L 61100 Powder, mg/L
Basal Media
1.Eagle, H. (1959) Science, 130, 432. a. Original Formula lists this component as NaH2PO4, 2H2O2 * Also known as Nicotindamide.
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NCTC - 1351-4 Medium
Base cat. no. Component Inorganic salts CaCl2 2H2O Fe(NO3)3 9H2O KCl MgSO4 7H2O NaCl NaHCO3 NaH2PO4 2H2O Other components Ethyl Alcohol D-Glucose D-Glucuronolactone Phenol Red (sodium salt) Sodium Acetate 3H2O Sodium Glucuronate H2O Tween 80 Coenzymes CoA (coenzyme A) DPN (diphosphopyridine nucleotide) FAD (flavin adenine dinucleotide) TPN (triphosphopyridine nucleotide monosodium) TPP (cocarboxylase) UTP (uridine triphosphate, sodium) Reducing Agents Ascorbic Acid Glutathione, monosodium Nucleic Acid Derivatives Deoxyadenosine Deoxyguanosine Deoxycytidine HCI 5-Methylcytosine Thymidine Amino Acids L-Alanine L--Amino-n-butyric Acid L-Arginine HCI L-Asparagine L-Aspartic Acid L-Cystine Amino acids, continued D-Glucosamine HCI L-Glutamic Acid L-Glutamine Glycine L-Histadine HCI H20
41350 1X liquid, mg/L 264.00 0.10 400.00 200.00 6800.00 2200.00 158.00 40.00 1000.00 1.80 20.00 50.00 1.80 12.50 2.50 7.00 100 1.00 1.00 1.00
Base cat. no. Component Hydoxy-L-Proline L-Isoleucine L-Leucine L-LysineHCI L-Methionine L-Ornithine HCI L-Phenylalanine L-Proline L-Serine L-Taurine L-Threonine L-Tryptophan L-Tryosine L-Valine Vitamins -Tocopherol Phosphate (disodium salt) d-Biotin Calciferol (Vitamin D2) D-Ca-Panthothenate Choline Chloride Folic Acid i-Inositol Menadione (Vitamin K) Niacin Niacinamide* Para-aminobenzoic Acid Pyridoxal HCI Pyridoxine HCI Riboflavin Thiamine HCI Vitamin A (crystallised alcohol) Vitamin B12
41350 1X liquid, mg/L 4.10 18.00 20.40 38.40 4.40 9.40 16.50 6.10 10.80 4.20 18.90 17.50 16.40 25.00 0.025 0.025 0.25 0.025 1.25 0.025 0.125 0.25 0.625 0.003 0.125 0.0625 0.0625 0.025 0.025 0.25 10.00
50.00 10.00 10.00 10.00 10.00 0.10 10.00 31.48 5.50 31.20 8.10 9.91 10.50 3.90 8.30 135.70 13.50 26.50
1. Evans, V.J. et al, (1956) Can. Res., 16,77. 2. Mcquilken, W. T. Et al (1957) J. Nat. Cancer Inst., 19,885. 3. Evans V.J ., et al, (1964) Exp. Cell res., 36, 439. 4. Tissue Culture Standards Committee, In Vitro (1970) 6.2, 93 * Also known as Nicotindamide.
74
RPMI 1640 Medium1

Base cat. no. Component Inorganic salts Ca(NO3)2 4H2O KCl MgSO4 (anhyd.) MgSO4 7H2O NaCl NaHCO3 Na2HPO4 (anhyd.) Other components D-Glucose Glutathione (reuduced) HEPES Phenol red Amino acids L-Alanine HCI L-Asparagine (free base) L-Aspartic acid L-Cystine 2HCl L-Glutamic acid L-Glutamine Glycine L-Histidine (free base) L-Hydroxyproline L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine (disodium salt) L-Valine Vitamins D-Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Para-aminobenzoic acid Pyridoxine HCl Riboflavin Thiamine HCl Vitamin B12 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 0.20 0.25 3.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 1.00 0.005 241.86 50.00 20.00 65.15 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 28.94 20.00 200.00 50.00 20.00 65.00 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 28.00 20.00 241.86 50.00 20.00 65.15 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 28.94 20.00 240.00 50.00 20.00 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 20.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 241.86 50.00 20.00 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 20.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 2000.00 1.00 1.00 5.00 2000.00 1.00 5958.00 5.00 2000.00 1.00 5.00 2000.00 1.00 4770.00 5.00 2000.00 1.00 5.00 2000.00 100.00 400.00 48.84 6000.00 2000.00 800.00 100.00 400.00 48.84 6000.00 2000.00 1,512.00 100.00 400.00 48.84 5500.00 800.00 100.00 400.00 100.00 6000.00 2000.00 800.00 100.00 400.00 100.00 6400.00 1000.00 800.00 100.00 400.00 100.00 6000.00 2000.00 800.00 100.00 400.00 100.00 6000.00 2000.00 800.00 11835 1X liquid, mg/L 11879 1X liquid, mg/L 13018 Powder, mg/L 21875 1X liquid, mg/L 22409 1X liquid, mg/L 27016 1X liquid, mg/L 31870 1X liquid, mg/L
1.00 5.00
Basal Media
1. Moore, G. E., .Gerner, R. E. And Franklin, H,. A. (1967) A. M. A., 199 519. * Also known as Nicotindamide.
75
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RPMI 1640 Medium1

Base cat. no. Component Inorganic salts Ca(NO3)2 4H2O KCl MgSO4 (anhyd.) MgSO4 7H2O NaCl NaHCO3 Na2HPO4 (anhyd.) Other components D-Glucose Glutathione (reuduced) HEPES Phenol red Amino acids L-Arginine HCI L-Asparagine (free base) L-Aspartic acid L-Cystine L-Cystine 2HCl L-Glutamic acid L-Alanyl-L-Glutamine L-Glutamine Glycine L-Histidine (free base) L-Hydroxyproline L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine (disodium salt) L-Valine Vitamins D-Biotin D-Ca pantothenate Choline chloride Folic acid i-Inositol Niacinamide* Para-aminobenzoic acid Pyridoxine HCl 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 0.20 0.30 3.00 1.00 35.00 1.00 1.00 1.00 0.20 0.25 3.00 1.00 35.00 1.00 1.00 1.00 240.00 50.00 20.00 50.00 20.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 50.00 20.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 50.00 20.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 65.15 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 28.94 20.00 240.00 50.00 20.00 50.00 20.00 300.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 50.00 20.00 446.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 240.00 50.00 20.00 50.00 20.00 446.00 10.00 15.00 20.00 50.00 50.00 40.00 15.00 15.00 20.00 30.00 20.00 5.00 20.00 20.00 2000.00 1.00 2000.00 1.00 5958.00 5.00 2000.00 1.00 4770.00 5.00 2000.00 1.00 5.00 2000.00 1.00 5958.00 5.00 2000.00 1.00 5.00 2000.00 1.00 5958.00 5.00 100.00 400.00 100.00 6000.00 2000.00 800.00 100.00 400.00 100.00 5500.00 2000.00 800.00 100.00 400.00 100.00 6000.00 850.00 800.00 100.00 400.00 48.84 6000.00 800.00 100.00 400.00 100.00 5500.00 2000.00 800.00 100.00 400.00 100.00 6000.00 2000.00 800.00 100.00 400.00 100.00 5500.00 2000.00 800.00 30244 1X liquid, mg/L 42401 1X liquid, mg/L 42402 1X liquid, mg/L 51800 Powder, mg/L 52400 1X liquid, mg/L 61870 1X liquid, mg/L 72400 1X liquid, mg/L
76
RPMI 1640 Medium1

Base cat. no. Component Vitamins, continued Riboflavin Thiamine HCl Vitamin B12 0.20 1.00 0.005 0.20 1.00 0.005 0.20 1.00 0.005 0.20 1.00 0.005 0.20 1.00 0.005 0.20 1.00 0.005 0.20 1.00 0.005 30244 1Xliquid, mg/L 42401 1X liquid, mg/L 42402 1X liquid, mg/L 51800 Powder, mg/L 52400 1X liquid, mg/L 61870 1X liquid, mg/L 72400 1X liquid, mg/L
1. Moore, G. E., .Gerner, R. E. And Franklin, H,. A. (1967) A. M. A., 199 519. * Also known as Nicotindamide.
3 Basal Media 77
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Advanced RPMI Medium 1640

Base cat. no. Component Inorganic salts Calcium nitrate (Ca(NO3)2 4H2O) Magnesium Sulfate (MgSO4-7H2O) Potassium Chloride (KCI) Sodium Bicarbonate (NaHCO3) Sodium Chloride (NaCI) Sodium Phosphate dibasic (Na2HPO4-7H2O) Zinc Sulphate (ZnSO47H2O) Other components D-Glucose (Dextrose) Ethanolamine Glutathione (reduced) Phenol red Sodium Pyruvate Amino acids Glycine L-Alanine L-Arginine L-Asparagine L-Aspartic Acid L-Cystine 2HCI L-Glutamic Acid L-Histidine L-Hydroxyproline L-Isoleucine L-Leucine L-Lysine hydrochloride L-Methionine L-Phenylalanine L-Proline L-Serine L-Theronine L-Tryptophan L-Tyrosine (disodium salt) L-Valine Vitamins Ascorbic Acid Phosphate Biotin Choline chloride D-Calcium pantothenate Folic acid i-Inositol 2.5 0.2 3 0.25 1 35 10 8.9 200 50 20 65 20 15 20 50 50 40 15 15 20 30 20 5 29 20 2000 1.9 1 5 110 100 100 400 2000 6000 1512 0.874 12633 1X liquid, mg/L Niacinamide Para-Aminobenzoic Acid Pyridoxine hydrochoride Riboflavin Thiamine hydrochoride Vitamin B12 Proteins AlbuMAX II Human Transferrin (Holo) Insulin Recombinant Full Chain Reducing Agents Glutathione, monosodium Trace Elements Ammonium Metavanadate Cupric Sulfate Manganous Chloride Sodium Selenite 0.0003 0.00125 0.00005 0.005 1 400 7.5 10 1 1 1 0.2 1 0.005
Waymouths Medium
Base cat. no. Component Inorganic salts CaCl2 H24O KCl KH2PO4O MgCl2 6H2O MgSO4 7H2O NaCl NaHCO3 Na2HPO4 (anhyd.) Other components D-Glucose Glutathione (reduced) Hypoxanthine (sodium salt) Phenol red Amino acids L-Arginine HCl L-Aspartic acid L-Cysteine HCl L-Cystine L-Glutamic acid L-Glutamine Glycine L-Histidine (free base) L-Isoleucine L-Leucine L-Lysine HCl L-Methionine L-Phenylalanine L-Proline L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins Ascorbic acid Biotin D-Ca pantothenate Choline Chloride Folic Acid i-Inositol Niacinamide* Pyridoxine HCl Riboflavin Thiamine HCl Vitamin B12 17.50 0.02 1.00 250.00 0.40 0.90 1.00 1.00 1.00 10.00 0.20 75.00 60.00 90.00 15.00 150.00 350.00 50.00 128.00 25.00 50.00 240.00 50.00 50.00 50.00 75.00 40.00 40.00 65.00 5000.00 15.00 25.00 10.00 120.00 150.00 80.00 240.00 200.00 6000.00 2240.00 300.00 31220 1X liquid, mg/L
1. Waymouth, C. (1968) Personal Communivation. * Also known as Nicotindamide.
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Williams Medium E 1
Base cat. no. Component Inorganic salts CaCl2 2H2O CuSO4 5H2O Fe(NO3)3 9H2O KCl MgSO4 7H2O MnCl2 4H2O NaCl NaHCO3 NaH2PO4 2H2O ZnSO4 7H2O Other components D-Glucose Glutathione Methyl Linoleate Phenol Red Sodium Pyruvate Amino acids L-Alanine L-Arginine L-Asparagine H2O L-Aspartic Acid L-Cysteine L-Cystine L-Ananyl-L-Glutamine L-Glutamic Acid Glycine L-Histidine L-Isoleucine L-Leucine L-Lysine HCla L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Vitamins Ascorbic Acid Biotin D-Ca Pantothenate Choline Chloride Ergocalciferol Folic Acid i-Inositol 2.00 0.50 1.00 15.00 0.10 1.00 2.00 2.00 0.50 1.00 15.00 0.10 1.00 2.00 90.00 50.00 20.00 30.00 40.00 20.00 50.00 50.00 15.00 50.00 75.00 87.00 15.00 25.00 30.00 10.00 40.00 10.00 35.00 50.00 90.00 50.00 20.00 30.00 40.00 20.00 434.00 50.00 50.00 15.00 50.00 75.00 87.00 15.00 25.00 30.00 10.00 40.00 10.00 35.00 50.00 2000.00 0.05 0.03 10.00 25.00 2000.00 0.05 0.03 10.00 25.00 264.00 0.0001 0.0001 400.00 200.00 0.0001 6800.00 2200.00 158.00 0.0002 264.00 0.0001 0.0001 400.00 200.00 0.0001 6800.00 2200.00 158.00 0.0002 22551 1X liquid, mg/L 32551 1X liquid, mg/L Menadione Sodium Bisulfate Niacinamide* Pyridoxal HCl Riboflavin -Tocopherol Phosphate, Disodium Thiamine HCl Vitamin A Acetate Vitamin B12 0.01 1.00 1.00 0.10 0.01 1.00 0.10 0.20 0.01 1.00 1.00 0.10 0.01 1.00 0.10 0.20
1. Williams, G. M. And Gunn, J. M. (1974) Exp. Cell. Res., 89, 139. Supplement with 292.00 mg/L L-Glutamine (2mM) a. Original formula lists Lisine at 70.00 mg/L * Also known as Nicotindamide.
3 Basal Media 79
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Chapter 4 Serum-free and Specialty Media and Reagents

Summary of Serum-Free, Protein-free, and Chemically Defined Media . . . . . . . . . . . . . . . . . . . . . . 85 Summary of Specialty Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Serum-free Media and Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86 Differences Between Serum-free, Protein-free, and Chemically Defined Media . . . . . . . . . . . . . 86 Custom Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87 3D Matrices and Scaffolds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88 Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 CHO Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 Customized CHO Medium Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 DG44 Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 293 Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93 Virus Production Medium Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 Viral Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 Insect Cell Expression Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98 Customized Insect Medium Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 Insect Medium Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103 Hybridoma and Monoclonal Antibody Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 Customized Hybridoma Medium Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 Cellular Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109 Bronchial Epithelial Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109 Dermatology Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 Hematology-Oncology Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 Hematopoiesis Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 Hepatocyte Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 Human Mammary Epithelial Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 Neurobiology Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
continued
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Stem Cell Research Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 B-27 Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 N-2 Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Neurobasal Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Neurobasal-A Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 AIM V Media (research grade) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 AIM V Media (therapeutic grade) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118 MesenPRO RS Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118 StemPro hESC SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 StemPro-34 SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 DMEM/F-12 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 KnocKout Serum Replacement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 KnocKout DMEM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 Geneticin Selective Antibiotic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122 Blasticidin S HCI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 Hygromycin B . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 Fetal Bovine Serum, ES Cell-Qualified . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 CO2-Independent Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 Basic Fibroblast Growth Factor (bFGF), Human, Recombinant . . . . . . . . . . . . . . . . . . . . . . . 124 Insulin-like Growth Factor-I (IGF-I), Human, Recombinant . . . . . . . . . . . . . . . . . . . . . . . . . . 124 Platelet Derived Growth Factor BB (PDGF BB), Human, Recombinant . . . . . . . . . . . . . . . . . . 124 Vascular-Endothelial Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124 Cytogenetics Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126 MarrowMAX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126 AmnioMAX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127 KaryoMAX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127 Related Products for Cytogenetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128 Overview of Transfection, Selection, and Protein Delivery Methods . . . . . . . . . . . . . . . . . . . . . . 131 Transfection Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Lipofectamine 2000 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 Lipofectamine LTX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 FreeStyle MAX Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132 293fectin Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 Lipofectamine RNAiMAX Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 Cellfectin Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 PLUS Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 Lipofectamine Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 Lipofectamine 2000 Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
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Important Licensing Information: The products in this catalogue may covered by one or more Limited Use Label Licenses (see the Invitrogen website www .invitrogen .com) . By use of these products, you accept the terms and conditions of all applicable Limited Use Label Licenses . All products are for research use only, and are not intended for human or animal diagnostics, therapeutic, commercial, or other clinical uses unless otherwise specifically indicated in the applicable product Limited Use Label License(s) .
Summary of Serum-free, Protein-free, and Chemically Defined Media

ApplicationRecombinant protein production Mammalian Cells CHO, DG44 HeLa, HEK 293-F, HEK 293-H Insect Sf9, Sf21, Tn-368 BTI-TN-5B1-4 (High Five) Monoclonal antibody production Hybridomas, myelomas, NS0, NS1, Sp2 Media CD OptiCHO Medium, CD CHO Medium, CD CHO AGT, CHO-S-SFM II CD 293 Medium, CD 293 AGT, 293 SFM II , Protein Expression Medium (PEM) Sf-900 III, SFM Sf-900 II SFM Express Five SFM CD Hybridoma Medium, CD Hybridoma AGT Hybridoma-SFM PFHM-II Protein-free Hybridoma Medium
ApplicationVirus production VERO, BHK-21, HEp-2, COS-7L HEK 293-F, HEK 293-H, HeLa MDBK, MDCK, BHK-21, PK-15 PER .C6 ApplicationCellular studies Dermatology Hematologyoncology Hematopoiesis Keratinocytes Lymphocytes (human primary lymphoid cells and transformed lymphoid lines) Hematopoietic cells (bone marrow, peripheral blood, and neonatal cord blood) Peripheral macrophages and monocytes Vascular research Hepatic research Neurobiology Transgenic mice Endothelial cells Hepatocytes Neuronal cells Embryonic stem cells (mouse) Defined Keratinocyte-SFM (without BPE) Keratinocyte-SFM (with BPE) AIM V Media VP-SFM, VP-SFM AGT CD 293 Medium, CD 293 AGT 293 SFM II OptiPRO SFM, Opti-MEM Protein Expression Medium (PEM), Adenovirus Expression Medium (AEM)
4 Serum-free and Specialty Media and Reagents
StemPro-34 SFM Macrophage-SFM Human Endothelial-SFM Hepatozyme-SFM Neurobasal Media, B-27 supplements KnocKout Serum Replacement KnocKout DMEM
For additional information on applications of serum-free media, see Appendix I, starting on page 187 .
Summary of Specialty Media*

Cell type Amniocytes, chorionic villus cells Specialty media AmnioMAX-II Complete Medium AmnioMAX-C100 Complete Medium (basal medium and supplement) Application In vitro propagation of primary human amniotic fluid cells for karyotyping In vitro propagation of primary human amniotic fluid cells for karyotyping Supplement contains growth supplements (serum) and antibiotics Lymphocytes Bone marrow and peripheral blood Human mammary epithelial cells Bronchial epithelial cells PB-MAX Karyotyping Medium MarrowMAX Bone Marrow Medium HuMEC Medium LHC Medium Fully supplemented (including FBS) RPMI-based medium for short-term cultivation of peripheral blood lymphocytes A fully supplemented medium developed specifically to support bone marrow and peripheral blood cells in culture for in vitro cytogenetic analysis of hematological disease Optimized specifically for the growth of human mammary epithelial cells Engineered specifically for the growth of bronchial epithelial cells such as normal human bronchial epithelial cells and bovine bronchial epithelial cells without further supplements
* Specialty media are optimized and designed for specific applications and require serum supplementation . For additional information on applications of specialty media, see Appendix I, starting on page 196 .
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Serum-free Media and Reagents

For information on classifications, production, or preparation of these products, see page 7 .
Boost Productivity, Meet Regulatory Requirements, and Simplify Purification

Researchers who think downstream choose gibco products for serum-free cell culture . If youre working with cells that may ultimately be used in genomic, proteomic, biopharmaceutical, therapeutic, or diagnostic applications, your choice of culture media becomes more critical than ever . Gibco media meet pharmaceutical standards for quality, and are the most approved in the biopharmaceutical industry because they: Minimize variables Maximize yield Simplify purification Meet regulatory requirements Optimize results
Differences Between Serum-free, Protein-free, and Chemically Defined Media

Serum-free media
Gibco Serum-free Media do not require supplementation with serum, but may contain discrete proteins or bulk protein fractions .
Protein-free media
Gibco protein-free media contain no proteins, but may contain plant or yeast hydrolysates . Many are animal originfree .
N IMA A FREE L O RIGIN
Chemically defined media
Optimize serum-free cell culture with gibco products

Choose from chemically defined, protein-free, and serum-free formulations Minimize risk with animal originfree media and reagents Save time with preadapted cells and cell line development kits Use liquid or Advanced Granulation Technology (AGT) formats that are scalable to large-volume cultures
Gibco chemically defined media contain no proteins, hydrolysates, peptides, or components of unknown composition . These media are animal originfree, and all components have a known chemical structure . Completely defined system eliminates variability Consistent performance improves reproducibility Decrease possibility of contamination by adventitious agents Save time with simplified purification and downstream processing
Minimize risk with animal originfree media and reagents

Materials of animal origin have the potential to introduce adventitious agents into a cell culture system . At Life Technologiestm we have developed a wide range of cell culture products that are completely free of animal-origin materials, by taking multiple steps to ensure the quality, safety, consistency, and regulatory compliance of our products: Sourcing nonanimal origin raw materials whenever possible Substituting bioactive synthetic chemicals for their native animal-derived counterparts Ensuring solid documentation that details source, origin, and manufacturing process for all raw materials Using non-protein compounds instead of insulin and transferrin in protein-free and chemically defined media Gibco animal originfree products do not contain material directly derived from animal tissues, cells, or body fluids of higher eukaryotic organisms such as mammals (including humans), fish, birds, insects, etc . The term animal origin does not pertain to lower eukaryotic organisms such as the higher plants, fungi, protozoa, and algae, nor does it include prokaryotic organisms such as bacteria or blue-green algae .
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Custom Formulations
All serum-free formulations can be customized to meet your specifications . Our Custom Products Services group will work with you to determine composition, processing, packaging, labeling, shipping and storage conditions, and pricing for your formulation .
Limited-edition stocked customs

Some formulations or supplements either in development or that have been requested as a custom by several customers may be offered as limited-edition stocked customs . This means we will have some made and ready to ship at your request . To learn more about additional options currently available, go to www .invitrogen .com/stockedcustoms .
Production, Preparation, and Quality Control

All serum-free formulations are produced and prepared to the same quality standards that apply to all our gibco products . For information on medium production, preparation, product packaging, closures, quality control procedures, and storage and sterilization, see page 7 . For general outlines of methods of adaptation of cultures to serum-free conditions, direct and sequential (weaning), see Appendix I, page 187 .
4 Serum-free and Specialty Media and Reagents 87

3D Matrices and Scaffolds

Geltrex matrix and the AlgiMatrix 3D Culture System contribute to a broader product portfolio of gibco 3D culture systems, which were developed to meet the demand for 3D cell culture products in fields such as stem cell and cancer research, drug development, toxicology, and development and morphogenesis, as well as tissue and organ engineering .
Geltrex Reduced Growth Factor Basement Membrane Matrix

Geltrex matrix is a soluble form of reduced growth factor (RGF) basement membrane extract (BME) purified from continuous sheets of specialized extracellular matrix that form an interface between Engelbreth-HolmSwarm (EHS) tumor cells . The major components of Geltrex matrix include laminin, collagen IV, entactin, and heparin sulfate proteoglycan, which provide the foundation for three-dimensional (3D) culture studies . Like naturally occurring basement membrane, Geltrex reduced growth factor basement membrane not only supports cells and cell layers, but also plays an essential role in tissue organization that affects cell adhesion, migration, proliferation, and differentiation . It is easy to prepare suspensions using Geltrex matrix, and it offers, on average, a 2-fold reduction in several growth factors . As a result, analyses of specific growth factor effects in applications requiring basement membrane extract can be carried out in a controlled and reproducible way using Geltrex matrix . Recommended storage conditions: 20C to 80C .
AlgiMatrix 3D Culture System

The AlgiMatrix 3D Culture System is a user-friendly nonanimal-derived bioscaffold for three-dimensional (3D) cell culture . The AlgiMatrix 3D Culture System creates a cell culture model system that more closely reflects normal cell morphology and behavior, to meet researchers requirements for 3D cell culture in fields such as toxicology, drug development, cancer and stem cell research, development and morphogenesis, tissue and organ engineering, heart disease, and diabetes . The AlgiMatrix 3D Culture System is supplied in a 96-well format that fits well into the workflows of both lower- and higher-throughput users . Scientists can inoculate cells directly into the sterile microtiter plates preloaded with lyophilized alginate sponge, with no pretreatment or preparation required . The combination of convenience, high capacity, and microscopic visualization makes the AlgiMatrix 3D Culture System an ideal solution for broad basic research and drug discovery applications . Other commercially available 3D matricesincluding agar, collagen, and Matrigelare either difficult to use or are of animal origin, with significant lot-to-lot variability that can compromise performance . The AlgiMatrix sponge is formulated using pharmaceutical-grade raw material from brown seaweed . Its pure, chemically defined, and nontoxic macroporous structure yields improved lot-to-lot consistency, superior cell loading, and excellent nutrient delivery, without damage to cells . Recommended storage conditions: room temperature .
Quantity 1 ml 5 ml Cat. no. 12760-013 12760-021
Related products Page HuMEC-SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 KnocKout Serum Replacement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Phosphate-Buffered Saline (PBS) 7 .2 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . .125 TrypLE Express Stable Trypsin Replacement Enzyme without phenol red . . . . . . . . . . . . . . . . . . . . . .162 Versene 1:5,000 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .163 Visit us at www .invitrogen .com/3D-CellCulture to learn more about the Geltrex 3D Culture System and related products for 3D cell culture .
Quantity 1 96-well plate 5 96-well plates Cat. no. 12684-015 12684-031
Collagen I, Rat Tail

Collagen I, Rat Tail is the most common fibrillar collagen (90%) found in skin, bone, tendons and other connective tissues . It is supplied at 5 mg/ml in a 20 ml vial providing the flexibility to dilute to lower concentrations . The detailed gelling procedure forms a firm, clear gel for 3D applications . Recommended storage conditions: 20C to 80C, in the dark .
Related products Page Defined Keratinocyte-SFM System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .110 Hepatozyme-SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .113 KnocKout Serum Replacement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .120 Phosphate-Buffered Saline (PBS) 7 .2 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . .141 TrypLE Express Stable Trypsin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .160 Versene 1:5,000 (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .161 Visit www .invitrogen .com/3D-CellCulture to learn more about the AlgiMatrix 3D Culture System and related products for 3D cell culture .
Quantity 20 ml Cat. no. A1048301
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Expression Products
FreeStyle MAX CHO Expression System
CHO Expression Products

The FreeStyle MAX CHO Expression System is a completely animal originfree protein expression system that provides scalable large-scale transient transfection and protein production in suspension FreeStyle CHO-S cells . FreeStyle CHO-S cells are Chinese hamster ovary (CHO) cells that have been adapted to serum-free suspension culture in FreeStyle CHO Expression Medium . Transfection and expression experiments may be performed directly in FreeStyle CHO Expression Medium without the need to change medium . The complete FreeStyle MAX CHO Expression System provides enough reagents to perform 25 transfections and one control transfection in a 30 ml volume, but larger-volume transfections may be performed using simple scale-up of reagents . At scale, the FreeStyle MAX CHO Expression System can provide milligram quantities of proteins in days instead of months . The FreeStyle MAX CHO Expression System includes the following major components: FreeStyle MAX ReagentThis transfection reagent provides high transfection efficiency in suspension FreeStyle CHO-S cells . FreeStyle CHO-S cellsThis cell line is adapted to high-density, serum-free suspension culture in FreeStyle CHO Expression Medium and is capable of producing high levels of recombinant protein . CHO cells provide stable and accurate glycosylation of proteins for downstream preclinical and therapeutic applications . FreeStyle CHO Expression MediumThis is a defined, serum-free medium formulated specifically to allow growth and large-scale transfection of suspension FreeStyle CHO-S cells . OptiPRO Serum-free Medium to facilitate optimal formation of DNA-lipid complexes . Recommended storage conditions: FreeStyle CHO cells, liquid nitrogen; FreeStyle MAX Reagent, 4C, do not freeze; FreeStyle CHO Expression Medium and OptiPRO Serum-free Medium, 4C, in the dark; pCMVSPORT--gal control vector, 20C .
CD OptiCHO Medium
Chemically defined (CD) medium optimized for recombinant CHO cells . CD OptiCHO Medium is specifically designed to offer high performance and yield with recombinant CHO cells in a chemically defined environment . The protein-free, animal originfree formulation provides consistency and reliability and reduces the need to screen for adventitious agents . Optimized to support and maximize recombinant CHO cell growth and protein expression in suspension culture Provides up to 40% greater growth and production than existing media for selected cell lines and applications Truly chemically defined and protein-free formulation contains no hydrolysates, improving lot-to-lot consistency and reliability Recommended storage conditions: 2C to 8C, in the dark . Guaranteed stable for 12 months when properly stored .
Quantity 1,000 ml 6 1,000 ml Cat. no. 12681-011 12681-029
Serum-free and Specialty Media and Reagents
Quantity 1 kit
80
10 L Wave transfection 50 ml control transfection in a shake ask
56.4 51.2
Cat. no. K9000-20

Viable cell density (cells/ml)
1 107
400
300
Human IgG yield (mg/L)
80 80 80 80 80 80 80 80
1.5
70.1 67.5 60.1
66.6 62.2
200 1 106
CD OptiCHO (viable cell density) CD CHO (viable cell density) CD OptiCHO (EPO) CD CHO (EPO)
100
37.9 29.6
0
9.8 12.5
Time (days)
10
12
11
12
Days after transfection
Transient transfection using FreeStyle MAX Reagent with FreeStyle CHO cells in a 10 L Wave Bioreactor for antibody production.
Optimized growth of CHO cells in CD OptiCHO Medium. Recombinant CHO cells were seeded in 30 ml of CD OptiCHO Medium or CD CHO Medium at a viable cell density of 4 105 cells/ml in a 125 ml Corning shake flask . Cell density and viability were monitored daily using a Vi-CELL XR Cell Counter (Beckman Coulter) . Recombinant protein (EPO) expression level was monitored by ELISA .
EPO (mg/L)
89
CD CHO Medium (1X), liquid
CHO-S-SFM II
CHO-S-SFM II is a complete, serum-free, low-protein (less than 100 g/ ml) medium optimized for the growth and maintenance of Chinese hamster ovary (CHO) cells in suspension culture and for the production of recombinant proteins expressed by CHO cells .1,2 This ready-to-use medium is suitable for the suspension culturing of CHO and other cells in batch, continuous, and perfusion culture systems . Recommended storage conditions: 2C to 8C, in the dark . Notes: A CHO Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
CD CHO Medium is a chemically defined, protein-free medium optimized for the growth of Chinese hamster ovary (CHO) cells and expression of recombinant proteins in suspension culture . CD CHO Medium contains no proteins or peptide components of animal, plant, or synthetic origin . There are also no undefined lysates or hydrolysates in the formulation . CD CHO Medium is made without hypoxanthine and thymidine for use in dihydrofolate reductase (DHFR) amplified systems; for other applications, add 10 ml per liter of HT Supplement (Cat . no . 11067-030) prior to use Formulated without L-glutamine for increased stability; add 40 ml of 200 mM L-glutamine per 1,000 ml of medium prior to use Recommended storage conditions: 2C to 8C, in the dark . Notes: A CHO Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
References
1 . Tilkins, M .L ., Judd, D .A ., Weiss, S .A ., and Gorfien, S .F . (1992) FOCUS 14, 95 . 2 . Battista, P .J ., Tilkins, M .L ., Judd, D .A ., Godwin, G .P ., and Gorfien, S .F . (1994) American Biotechnology Laboratory 12:5, 64 .
Quantity 500 ml 1,000 ml
Contains no glutamine, hypoxanthine, thymidine, or phenol red .
Cat. no. 10743-011 10743-029 Quantity 500 ml 1,000 ml
Contains L-glutamine .
Cat. no. 12052-114 12052-098
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123 HT Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .107 CHO-S Cells adapted to CD CHO Medium . . . . . . . . . . . . . . . . . . . . . . . . . .178 DG44 Cells adapted to DG44 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .178
Quantity 500 ml
Contains L-glutamine, but no hypoxanthine or thymidine .
Cat. no. 31033-020
Customized CHO Medium Products

We offer several CHO formulations that are not routinely stocked in inventory, but are available on a make-to-order basis . We realize that not all needs are alike, so weve developed the capabilities to provide the desired configuration (formulation and packaging) of product type, as well as the operational systems that can provide these products in a timely fashion . Contact our Custom Products Services group for assistance in selecting the product that best meets your needs Customized products are available in a wide array of packaging, from standard PET bottles (100 ml, 500 ml, and 1,000 ml) to custom media bags (1 L, 5 L, 10 L, and 200 L) . We offer the following on a custom basis: Adherent CHO-SFM: designed for anchorage-dependent CHO systems . CHO-III-PFM: devoid of animal-sourced raw materials, this essentially protein-free medium contains a vegetable hydrolysate for added performance . Notes: A CHO-III-PFM Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative . Please note that variations of the above products, such classical media as DMEM/F-12, Nutrient Mixture F-12, and MCDB 301, as well as your own in-house developed formulations, can be tailored to your specific selection/amplification system (DHFR, glutamine synthetase, neomycin resistance, etc .) .
CD CHO AGT
CD CHO AGT is a chemically defined, protein-free medium in a new easy-to-use granular format . CD CHO AGT is optimized for the growth of Chinese hamster ovary (CHO) cells and expression of recombinant proteins in suspension culture . Recommended storage conditions: 2C to 8C, in the dark .
Quantity 11L 1 10 L Cat. no. 12490-017 12490-025
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123 HT Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .107 CHO-S Cells adapted to CD CHO Medium . . . . . . . . . . . . . . . . . . . . . . . . . .178 DG44 Cells adapted to DG44 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .178
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CHO CD EfficientFeed Kit

The CHO CD EfficientFeed Kit is a system designed to help you more quickly and efficiently identify a feed supplement formulation that will improve fed-batch productivity 2- to 5-fold versus batch conditions, as well as identify a feed supplement formulation that will help develop bioreactor cell seed stocks more quickly . The kit contains feed supplements A and B, based on successful chemically-defined media formulations you may be using or considering for your batch culture medium (CD OptiCHO Medium or CD CHO Medium .) Recommended storage conditions: 2C to 8C, in the dark .
DG44 Expression Products

DG44 Cells and Media Kit
The DG44 Cells and Media Kit are optimized for the growth of dihydrofolate reductasedeficient (DHFR) Chinese hamster ovary (CHO) cells and expression of recombinant proteins in suspension culture . The kit provides CD DG44 Medium, a chemically defined, protein-free medium specifically designed to enable optimal growth and expression of DG44 cells in suspension culture, and DG44 cells preadapted to CD DG44 Medium . Cells are selected for superior cell growth and transfection efficiencies . In addition, the DG44 Cells and Media Kit includes L-glutamine and Pluronic F-68 for optimal growth and protein expression of DG44 cells in a protein-free, chemically defined environment . The cells are banked under cGMP conditions and with full documentation available with commercial use license . Recommended storage conditions: CD DG44 Medium, 2C to 8C, in the dark; DG44 cells, in liquid nitrogen; L-glutamine, 5C to 20C; Pluronic F-68, 15C to 30C . Notes: A CHO Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contac ting I nvitrogen Technical Suppor t or your local Sales Representative .
Pluronic is a registered trademark of BASF Corporation .
Product CHO CD EfficientFeed Kit Kit components also available separately CHO CD EfficientFeed A CHO CD EfficientFeed B 1000ml 1000ml A10234-01 A10240-01 Quantity 1 kit Cat. no. A10241-01
Quantity 1 Kit Cat. no. A1100-001
91
Chapter 4 Serum-free and Specialty Media and Reagents Ordering Information

Product OptiCHO Protein Express Kit Kit components, also available separately pOptiVEC-TOPO TA Cloning Kit CD DG44 Medium FreeStyle MAX Reagent OptiPRO SFM CD OptiCHO Medium Pluronic F-68, 10% L-glutamine-200 mM (100X) 1 kit 1,000 ml 1 ml 100 ml 1,000 ml 100 ml 100 ml 12744-017 12610-010 16447-100 12309-050 12681-011 24040-032 25030-081 Quantity 1 kit Cat. no. A1099901
OptiCHO Protein Express Kit

The OptiCHO Protein Express Kit is designed for faster serum-free cell line development and cloning for single or two sub unit protein expression in suspension culture in dihydrofolate reductase-deficient (DHFR-) Chinese hamster ovary (CHO) DG44 cells created under cGMP condiitons .The Kit provides a complete workflow protocol and manual information provided on a reuseable USB flash drive, as well as the necessary components for efficient transfection and cloning of DHFR-DG44 cells in a serum-free environments and subsequent development of a stable cell line for protein production . The Kit contains: Two expression vectors for light and heavy chains-the pOptiVECTOPO bicistronic cloning vector and pcDNA 3 .3 TOPO cloning vector, the vectors are TOPO adapted to enable cloning of a PCR product containing your gene of interest with >85% efficiency . FreeStyle MAX Reagentan animal origin free transfection reagent offering high transfection efficiencies and consistent results . CD OptiCHO Medium a chemically defined, protein free medium (without hypoxanthine or thymidine) specially designed for high-density suspension of culture of recombinant CHO DG44 cells and secreted protein yield . CD DG44 Mediuma chemically defined, protein free medium supplemented with hypoxanthine and thymidine to support optimal growth of DG44 cells in suspension culture . DG44 cells (banked under cGMP conditions and with full documentation available with commercial use license) preadapted to CD DG44and selected for superior cell growth and transfection efficiencies .
92
293 Expression Products

CD 293 Medium (1X), liquid
CD 293 Medium is a chemically defined, protein-free medium optimized for the growth of suspension cultures of 293 cells . CD 293 Medium contains no proteins or peptides of animal, plant, or synthetic origin . There are also no undefined lysates or hydrolysates in the formulation . Formulated without L-glutamine for increased stability; add 20 ml of 200 mM L-glutamine or 20 ml of GlutaMAX-I Supplement per 1,000 ml of medium prior to use . Notes: A 293 Medium Master file has been submitted to the FDA . If you wish to cross-reference a Master file for CD 293 Medium, please contact Invitrogen Technical Support or your local Sales Representative . Recommended storage conditions: 2C to 8C, in the dark .
293 SFM II (1X), liquid

293 SFM II is a serum-free, low-protein (<10 g/ml) formulation optimized for high-density suspension culture of 293 cells .1 (It is not suitable for adherent 293 cell cultures .) This product replaces 293 SFM (Cat . no . 10970-010) . 293 SFM II is animal originfree . Formulated without L-glutamine for increased stability; add 20 ml of 200 mM L-glutamine or 20 ml of GlutaMAX-I Supplement per 1,000 ml of medium prior to use . Notes: A 293 Medium Master file has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative . Recommended storage conditions: 2C to 8C, in the dark .
Reference
1 . Epstein, D .A ., Godwin, G .P ., Gruber, D .F ., and Grefrath, P .L . (1999) FOCUS 21, 22 .
Quantity 1,000 ml Cat. no. 11913-019
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123 293-F Cells adapted to CD 293 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . .177 293-H Cells adapted to CD 293 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . .177
CD 293 AGT
CD 293 AGT is a chemically defined, protein-free medium in a new easy-to-use granular format . CD 293 AGT is optimized for the growth of suspension cultures of 293 cells . Recommended storage conditions: 2C to 8C, in the dark .
FreeStyle 293 Expression Medium

FreeStyle 293 Expression Medium is a chemically defined, protein-free medium specifically developed for the ability to support the growth and transfection of 293-F cells under suspension-type culture conditions . The medium is a complete, ready-to-use medium that has been supplemented with GlutaMAX-I Supplement and is animal originfree . FreeStyle 293 Expression Medium is able to save significant time and costs associated with adaptation of cell cultures to serum-free conditions . Recommended storage conditions: 2C to 8C, in the dark .
References
1 . Graham, F .L ., Smiley, J ., Russell, W .C ., and Nairn, R . (1977) J. Gen. Virol. 36, 59 . 2 . Harrison, T ., Graham, F ., and Williams, J . (1977) Virology 77, 319 . 3 . Ciccarone, V ., Chu, Y ., Schifferli, K ., Pichet, J .P ., Hawley-Nelson, P ., Evans, K ., Roy, L ., and Bennett, S . (1999) FOCUS 21, 54 . 4 . Epstein, D .A ., Godwin, G .P ., Gruber, D .F ., and Grefrath, P .I . (1999) FOCUS 21, 22 .
Quantity 1,000 ml 6 1,000 ml (case) 8 L fill in 20 L Wave Cellbag Cat. no. 12338-018 12338-026 0070049BB
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Virus Production Medium Products
FreeStyle 293 Expression System

The FreeStyle 293 Expression System simplifies and streamlines production of protein in 293 cells . Using the FreeStyle 293 Expression System, you can transfect 293 cells in suspension, eliminating the need to carry multiple flasks of adherent cells when large numbers of cells are required . The FreeStyle 293 Expression System offers the following key components to optimize transfection and expression in 293 cells: 293fectin Reagenta transfection reagent for the highest transfection efficiency in suspension 293 cells Gibco FreeStyle 293 Expression Mediuma chemically defined, protein-free medium specifically designed to enable efficient transfection and optimal growth of 293 cells in suspension FreeStyle 293-F cellsa clone of 293 cells with superior handling properties in FreeStyle 293 Expression Medium FreeStyle 293-F cells, 293fectin Reagent, and gibco FreeStyle 293 Expression Medium work together to give you the most robust production of bioactive protein possible . In addition, the FreeStyle 293 Expression System provides Gibco Opti-MEM I for improved DNA-lipid complex formation and a -galactosidase expression plasmid to test transfection efficiency . The FreeStyle 293 Expression System contains enough reagents to perform twenty-five 30 ml transfections, but is designed to be scalable so that other transfection volumes can easily be used if required . Recommended storage conditions: 293fectin, gibco FreeStyle 293 Expression Medium, and Gibco Opti-MEM, 4C; pCMVSPORT--gal, 20C; FreeStyle 293-F cells, in liquid nitrogen; gibco FreeStyle 293 Expression Medium, in the dark .
OptiPRO SFM (1X), liquid
OptiPRO SFM is an animal originfree serum-free medium designed for growth of several kidney-derived cell lines including MDCK, MDBK, VERO, BHK-21, and PK-15, which are important for virus and recombinant protein production . Recommended storage conditions: 2C to 8C, in the dark . Notes: An OptiPRO SFM Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
This product is the subject of US Patent No . 6,767,741 .
Opti-MEM I Reduced-Serum Medium (1X), liquid (with GlutaMAX-I)

A GlutaMAX-I medium . Recommended storage conditions: 2C to 8C, in the dark . Intended use: In vitro diagnostic (IVD) .
Product FreeStyle 293 Expression Kit GIBCO FreeStyle 293 Expression Medium 293fectin Transfection Reagent FreeStyle 293-F Cells Quantity 1 kit 1,000 ml 1 ml 1 107 cells Cat. no. K9000-01 12338-018 12347-019 R790-07
Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, trace elements, growth factors, and phenol red reduced to 1 .1 mg/L .
Opti-MEM I Reduced-Serum Medium (1X), liquid

Recommended storage conditions: 2C to 8C, in the dark . Intended use: In vitro diagnostic (IVD) .
Customized 293 Expression Medium Products

293 Expression Medium products and your in-house formulations are available on a make-to-order basis . We realize that not all needs are alike, so weve developed the capabilities to provide the desired configuration (formulation and packaging) of product type, as well as the operational systems that can provide these products in a timely fashion . Please contact our Custom Products Services group for assistance in selecting the products and packaging that best meet your requirements .
Contains HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1 .1 mg/L .
94
Opti-MEM I Reduced-Serum Medium (1X), liquid (without phenol red)

Recommended storage conditions: 2C to 8C, in the dark .
VP-SFM (1X), liquid
Contains HEPES buffer, 2,400 mg/L sodium bicarbonate, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, and growth factors, but no phenol red .

A modification of MEM (Eagles) . Recommended storage conditions: 2C to 8C, in the dark . Intended use: In vitro diagnostic (IVD) . See page 84 for procedure to prepare powdered Opti-MEM I, 10X .
VP-SFM is a serum-free, ultralow-protein (5 g/ml) medium containing no components of animal or human origin . It is designed for the growth of VERO cells for virus production .1 This medium is suitable for the growth of COS-7, MDCK, BHK-21 (suspension culture), and HEp-2 cells, and for recombinant protein production . Formulated without L-glutamine for increased stability; add 20 ml of 200 mM L-glutamine or 20 ml of GlutaMAX-I Supplement per 1,000 ml of medium prior to use . Recommended storage conditions: 2C to 8C, in the dark . Notes: A VP-SFM Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
Reference
1 . Price, P . and Evege, E . (1997) FOCUS 19, 67 .
This product is the subject of US Patent No . 6,103,529 .
Quantity 10 1 L 1 10 L Cat. no. 22600-050 22600-134
Contains HEPES buffer, hypoxanthine, thymidine, sodium pyruvate, L-glutamine, trace elements, growth factors, and phenol red reduced to 1 .1 mg/L, but no sodium bicarbonate .
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123
VP-SFM AGT
VP-SFM AGT is a serum-free, ultralow-protein (5 g/ml) medium in a new easy-to-use granular format . VP-SFM AGT is designed for the growth of VERO cells for virus production . It is also suitable for the growth of COS-7, MDCK, BHK-21, and HEp-2 cells and for recombinant protein production . Recommended storage conditions: 2C to 8C, in the dark .
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123 Distilled Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .161
95
Protein Expression Medium (PEM)
CD 293 Medium (1X), liquid
Efficient production of recombinant proteins in PER .C6 cells Supports the large-scale, high-density culture of transfected PER .C6 cells in shake flasks, roller bottles, and bioreactors Formulated without L-glutamine for maximum stability Supports parental PER .C6, HEK 293, and HeLa cell growth in suspension culture, as well as transfected PER .C6 cells for protein expression applications Recommended storage conditions: 2C to 8C, in the dark .
Contains no components directly derived from human or animal sources and one low-level, recombinant protein, minimizing the opportunity to introduce adventitious agents .
CD 293 Medium is a chemically defined, protein-free medium optimized for the growth of suspension cultures of 293 cells . CD 293 Medium contains no proteins or peptides of animal, plant, or synthetic origin . There are also no undefined lysates or hydrolysates in the formulation . Formulated without L-glutamine for increased stability; add 20 ml of 200 mM L-glutamine or 20 ml of GlutaMAX-I Supplement per 1,000 ml of medium prior to use . Recommended storage conditions: 2C to 8C, in the dark . Notes: A 293 Medium Master file has been submitted to the FDA . If you wish to cross-reference a Master file for CD 293 Medium, please contact Invitrogen Technical Support or your local Sales Representative .
Adenovirus Expression Medium (AEM)
Viable cell density (vc/ml)
1 106
400
200
1 105 0 1 2 3 4 5 6 7 8 9

PER .C6 is a registered trademark of Crucell Holland B .V .
Time (days)
PER.C6 EpCAM cell growth and IgG production in PEM, VPRO, and AEM. PER .C6 EpCAM cells were adapted from DMEM/F-12 into gibco PEM, SAFC/ Sigma EX-CELL VPRO Serum-free Medium for Retinoblast Cells, and gibco AEM over several passages until stable cultures were achieved . Data represent growth experiments performed in triplicate for each condition . Cells grown in AEM and VPRO media were seeded at 3 105 vc/ml into 250 ml shake flasks with a working volume of 40 ml; cells grown in PEM were seeded at 1 105 vc/ml into 250 ml shake flasks with a working volume of 40 ml, as PEM supports more optimized conditions with a lower seeding density and extended growth .
96
IgG (mg/L)
Adenovirus Expression Medium (AEM) is recommended for the growth of PER .C6 cells in suspension and for the production of viruses and viral vectors in these cells . It is an animal originfree cell culture medium without components directly derived from human or animal sources . Each lot of AEM is performance tested for cell growth with PER .C6 cells, and AEM has been qualified to produce adenovirus and influenza viruses . It also supports HEK 293 cell growth . Glutamine or GlutaMAX-I supplementation is required . Recommended storage conditions: 2C to 8C, in the dark . Notes: An Adenovirus Expression Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
1 107
PEM cell density VPRO cell density AEM cell density PEM IgG VPRO IgG AEM IgG
600
CD 293 AGT
Customized Virus Production Medium Products

Please note that the virus production products and your in-house formulations are available on a make-to-order basis . We realize that not all needs are alike, so weve developed the capabilities to provide the desired configuration (formulation and packaging) of product type, as well as the operational systems that can provide these products in a timely fashion . Please contact our Custom Products Services group for assistance in selecting the products and packaging that best meet your requirements .
CD 293 AGT is a chemically defined, protein-free medium in a new easy-to-use granular format . CD 293 AGT is optimized for the growth of suspension cultures of 293 cells . Recommended storage conditions: 2C to 8C, in the dark .
Viral Expression Products

Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123 Distilled Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .163 293-F Cells adapted to CD 293 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . .161 293-H Cells adapted to CD 293 Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . .177
ViraPower Adenoviral Expression System
The ViraPower Adenoviral Expression System has improved the process of making an adenoviral construct . Get clones with less effort and go directly to your expression experiments .
293 SFM II (1X), liquid
Product pAd/BLOCK-iT Adenoviral RNAi Vector BLOCK-iT U6 RNAi Entry Vector Kit pAd/PL-DEST Gateway Vector 293A Cell Line
* Kit contains Gateway DEST Vector and 293A Cell Line .
293 SFM II is an animal originfree, serum-free, low-protein (<10 g/ml) formulation optimized for high-density suspension culture of 293 cells .1 (It is not suitable for adherent 293 cell cultures .) This product replaces 293 SFM (Cat . no . 10970-010) . It is formulated without L-glutamine for increased stability . Add 20 ml of 200 mM L-glutamine or 20 ml of GlutaMAX-I Supplement per 1,000 ml of medium prior to use . Recommended storage conditions: 2C to 8C, in the dark . Notes: A 293 Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
Quantity 6g 1 kit 6 g 3 106 cells
Cat. no. K4930-00 V492-20 K4945-00 V494-20 R705-07
ViraPower Adenoviral Gateway Expression Kit* 1 kit
For more information on how ViraPower adenoviral expression and Gateway technology work, visit our website at www .invitrogen .com .
Reference
1 . Epstein, D .A ., Godwin, G .P ., Gruber, D .F ., and Grefrath, P .L . (1999) FOCUS 21, 22 .
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Insect Cell Expression Products
Overview of Stable Insect Expression Systems
Stable insect expression systems combine the advantages of both baculovirus and mammalian expression . Stable insect systems are simple to use with straightforward techniques for transfection and selection similar to those used with mammalian expression systems . At the same time, stable insect systems often achieve higher levels of expression than mammalian systems and are particularly useful for production of secreted proteins . Like baculovirus systems, high levels of expression are attainable using stable insect expression systems . An added benefit is the reliability and reproducibility of expression levels . This differs from baculovirus expression levels that can vary from experiment to experiment . Once a stable cell line is produced, the expression levels are maintained over time . In addition, stable insect expression systems produce protein from healthy, logarithmically growing cells, resulting in high-quality protein . The InsectSelect System is a stable insect expression system designed for continuous, non-lytic expression using Sf9, Sf21, or High Five cells . Stable expression with the InsectSelect system can be achieved in just three simple steps: 1 . Clone a gene of interest into an InsectSelect vector . 2 . Transfect insect cells . 3 . Select stable clones . The InsectSelect System uses blasticidin or Zeocin selection for efficient selection of stable clones . With blasticidin, clones can be established in as little as one week . The protein yield often depends on the nature of the gene . Yields of 110 g/ml can be expected but are not guaranteed .
Overview of Baculovirus Expression Systems

Expression in insect cells offers significant advantages, including high expression levels, ease of scale-up, production of proteins with posttranslational modifications, and simplified cell growth . Insect cells do not require CO2 for growth and can be readily adapted to high-density suspension culture for large-scale expression . Many of the posttranslational modification pathways present in mammalian systems are also utilized in insect cells, allowing the production of recombinant protein that is antigenically, immunogenically, and functionally similar to the native mammalian protein . Baculovirus expression systems are powerful and versatile systems for high-level recombinant protein expression in insect cells . Expression levels up to 500 mg/L have been reported using the baculovirus expression system, making it an ideal system for high-level expression . The BaculoDirect Baculovirus Expression System is the fastest and easiest method available for generating recombinant baculo virus . Purified virus can be isolated within one week without plaque purification . The protein yield often depends on the nature of the gene . Yields of 25250 g/ml can be expected but are not guaranteed . The BaculoDirect Linear DNA includes attR sites for rapid and efficient recombinational cloning with a Gateway entry clone . The resulting recombinant DNA can be taken directly from the LR reaction mix and used to transfect insect cells, saving a significant amount of time . Purified virus can be isolated within one week . The reduction of hands-on time makes the BaculoDirect system ideal for high-throughput expression . The Bac-to-Bac Baculovirus Expression System uses a unique bacmid shuttle vector that recombines with an expression cassette by site-specific transposition to create an expression bacmid . The expression bacmid is then transfected into insect cells to generate recombinant baculovirus . The positive expression bacmid is selected by a simple blue/white screening in E. coli . Once confirmed, it is transfected into insect cells to generate recombinant baculovirus . No tedious plaque purification is needed . The Bac-N-Blue Baculovirus Expression System has been used for over a decade to produce high levels of recombinant proteins . Improvements to the AcMNPV DNA, higher-expressing cell lines, and small vectors have made this system faster and easier to use .
Insect Expression Systems

Fusion partner System BaculoDirect Bac-to-Bac Bac-N-Blue Host Sf9, Sf21, or High Five Sf9, Sf21, or High Five Sf9, Sf21, or High Five Position C-term N-term N-term C-term Purification 6xHis 6xHis 6xHis Epitope V5 NA Xpress V5 Promoter Polyhedrin Polyhedrin or p10 Polyhedrin Advantage Easiest method of baculovirus production, ideal for HTP Rapid baculovirus production for small-scale expression High-level expression, well-characterized system
98
Sf-900 III SFM
Sf-900 II SFM
Sf-900 II SFM is a protein-free insect cell culture medium optimized for the growth and maintenance of Spodoptera frugiperda (Sf9 and Sf21) cells and for large-scale production of recombinant proteins expressed using the baculovirus expression vector system (BEVS) . This medium is suitable for suspension and monolayer culture methods and supports growth of other lepidopteran cell lines .1 Notes: An Sf-900 II SFM Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
Sf-900 III SFM is a protein-free, animal originfree medium that provides consistency and reliability in cell growth and protein expression . This product is formulated with one-third of the hydrolysate found in Sf-900 ll . Because this medium does not include animal-derived components, the need to screen for adventitious agents is reduced . Animal originfree medium with improved consistency in insect cell culture Optimized to support and maximize Sf9 and Sf21 cell growth and recombinant gene expression in suspension culture using both baculovirus and stable insect expression systems Supports Sf9 cell growth to densities of 10 to 14 106 cells/ml Protein-free, low-hydrolysate formulation improves lot-to-lot consistency and reliability Recommended storage conditions: 2C to 8C, in the dark . Guaranteed stable for 12 months when properly stored .
Reference
1 . Godwin, G .P . and Whitford, W . (1993) FOCUS 15, 44 .
Sf-900 II SFM (1X), liquid

Insect cell culture tested . Recommended storage conditions: 2C to 8C, in the dark .
Quantity 500 ml 1,000 ml 10 L Cat. no. 12658-019 12658-027 12658-035
Quantity 500 ml 1,000 ml 6 1,000 ml (case) 5L 20 L 10 L fill in 20 L Wave Cellbag
Cat. no. 10902-096 10902-088 10902-104 10902-161 10902-187 0070037BA
Contains no components of animal origin, facilitating the regulatory process .
16 106 14 106 12 10
6
Cells/ml
10 106 8 106 6 10
6
Related products Page Sf21 Cells adapted to Sf-900 II SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .178 Sf9 Cells adapted to Sf-900 II SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .177
Sf-900 II SFM (1X), liquid (without Met and Cystine)

Sf-900 II, lot #1
4 106 2 106 0 Day 4 Day 5
Sf-900 II, lot #2 Sf-900 III, lot #1 Sf-900 III, lot #2 Sf-900 III, lot #3
Quantity 500 ml
Contains L-glutamine, but no L-methionine or L-cystine .
Day 6
Day 7
Cat. no. 21012-026
Sf9 cell growth in Sf-900 II SFM and Sf-900 III SFM at passage 6. Sf9 cells were maintained in Sf-900 II SFM or Sf-900 III SFM media . After 6 passages in the test media, cell growth curves were determined by seeding 30 ml cultures in 125 ml shake flasks at 500,000 cells/ml, and counting viable cell number on each day post-seed . Data points represent the averages of 3 replicate flasks, and the error bars denote one standard deviation .
Related products Page Sf21 Cells adapted to Sf-900 II SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175 Sf9 Cells adapted to Sf-900 II SFM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .175
Sf-900 Medium (1.3X), liquid

99
Chapter 4 Serum-free and Specialty Media and Reagents Ordering Information

Quantity 100 ml
Cat. no. 10967-032

Schneiders Drosophila Medium (1X), liquid

Modified and insect cell culture tested . Recommended storage conditions: 2C to 8C .
Contains 3,330 mg/L lactalbumin hydrolysate, 3,330 mg/L yeastolate, and L-glutamine, but no insect hemolymph .

Express Five SFM (1X), liquid

Express Five SFM is a protein-free insect cell culture medium optimized to support maximum cell growth and recombinant gene expression by the baculovirus expression vector system (BEVS) in the BTI-TN-5B1-4 (High Five) Trichoplusia ni cell line . This unique medium eliminates problems associated with the culturing of the High Five cell line . It is optimized for serum-free growth in monolayer and suspension culture, boosting insect cell growth to densities exceeding 5 106 cells/ml and population doubling times of 16 to 30 hours . It demonstrates expression of recombinant proteins that is equal or superior to that of other serum-free formulations . It is formulated without L-glutamine for increased stability . Add 45 ml of 200 mM L-glutamine per 500 ml medium prior to use . Insect cell culture tested . Recommended storage conditions: 2C to 8C, in the dark .
Contains 6,660 mg/L lactalbumin hydrolysate, 6,660 mg/L yeastolate, and L-glutamine .
Graces Insect Cell Culture Medium, unsupplemented (1X), liquid

Quantity 500 ml
Contains L-glutamine, but no L-methionine or insect hemolymph .
Reference
1 . Godwin, G .P ., Danner, D ., and Gorfien, S .F . (1995) FOCUS 17, 58 .
Cat. no. 11595-030
IPL-41 Insect Medium (1X), liquid

Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149
Contains L-glutamine, calcium chloride, and sodium bicarbonate, but no tryptose phosphate broth .
100
Customized Insect Medium Products

We will customize the Insect Cell Expressions products formulations to best suit your systems needs . Please contact our Custom Products Services group for assistance in selecting the products and packaging that best meet your requirements Please note that Sf900 II Serum-free Medium has been found to be useful in insect cell culture systems other than those employing Sf9 or Sf21 lines . .
Insect Cell Culture Media Formulations

Base cat. no. Component Inorganic salts CaCl2 CuCl22H2O CoCl26H2O FeSO47H2O KCl KH2PO4 MgCl2 MgCl4H2O MgCl26H2O MgSO4 MgSO47H2O MnCl24H2O NaCl NaHCO3 NaH2PO4H2O NaH2PO42H2O NaH2PO47H2O Na2HPO4 (NH4)6(Mo7O244H2O) ZnCl2 Other components D-Fructose Fumaric acid D-Glucose -Ketoglutaric acid Malic acid Maltose Succinic acid Sucrose Trehalose2H2O N2 Amine YT Amino acids L-Alanine -Alanine 225 .00 200 .00 225 .00 200 .00 225 .00 200 .00 450 .00 400 .00 300 .00 500 .00 400 .00 55 .00 700 .00 370 .00 670 .00 60 .00 26,680 .00 400 .00 55 .00 700 .00 370 .00 670 .00 60 .00 26,680 .00 400 .00 55 .00 700 .00 370 .00 670 .00 60 .00 26,680 .00 800 .00 110 .00 1,400 .00 740 .00 1,340 .00 120 .00 53,360 .00 4 .40 2,500 .00 29 .60 53 .60 1,000 .00 4 .80 1,650 .00 100 .00 2,000 .00 200 .00 100 .00 2,000 .00 750 .00 4,100 .00** 1,070 .00 1,358 .00 1,013 .00 500 .00 2,800 .00 1,070 .00 1,358 .00 350 .00 1,013 .00 750 .00 4,100 .00 2,280 .00 2,780 .00 350 .00 1,013 .00 1,000 .00 0 .20 5,600 .00 2,138 .64 2,715 .34 700 .00 2,026 .00 500 .00 0 .20 0 .05 0 .55 1,200 .00 0 .02 918 .00 0 .02 2,850 .00 350 .00 1,160 .00 0 .04 0 .04 1,600 .00 450 .00 3700 .00 2,100 .00 400 .00 1321 .00 794 .00 794 .00 11300 Graces powder, mg/L
* *
11605 Graces supplemented 1X liquid, mg/L

*
11595 Graces unsupplemented 1X liquid, mg/L
11667 Graces 2X liquid, mg/L

*
11405 IPL-41L 1X liquid, mg/L
21720 Schneiders 1X liquid
* Grace, T .D . (1962) Nature 195, 788 . Schneider, Imogene (1964) Exp. Zool. 156:1, 91 . Revision of original formulation, Schneider, Imogene (1965) personal communications . Original formulations list Na2HPO 4 at 700 .00 mg/L . Revised formulation believed by author to possess better buffering capacity and better nutritive properties . ** Increased from 2,240 .00 mg/L upon recommendation of Dr . Vaughn .
continued
101
Insect Cell Culture Media Formulations, continued

Base cat. no. Component Amino acids, continued L-Arginine L-ArginineHCl L-Asparagine L-Aspartic acid L-Cysteine L-Cystine L-Cystine2HCl L-Cystine2Na L-Glutamic acid L-Glutamine Glycine L-Histidine L-Histidine HClH2O L-Hydroxyproline L-Isoleucine L-Leucine L-LysineHCl L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Tyrosine2Na L-Valine Vitamins Para-aminobenzoic acid Biotin D-Calcium pantothenate Choline chloride Folic acid i-Inositol Niacin Pyridoxine HCl Riboflavin Thiamine HCl Vitamin B12 Other components Lactalbumin Hydrolysate Yeastolate 3,303 .00 3,330 .00 6,660 .00 6,660 .00 2,000 .00 0 .02 0 .01 0 .02 0 .20 0 .02 0 .02 0 .02 0 .02 0 .02 0 .02 0 .02 0 .01 0 .02 0 .20 0 .02 0 .02 0 .02 0 .02 0 .02 0 .02 0 .02 0 .01 0 .02 0 .20 0 .02 0 .02 0 .02 0 .02 0 .02 0 .02 0 .04 0 .02 0 .04 0 .40 0 .04 0 .04 0 .04 0 .04 0 .04 0 .04 0 .32 0 .16 0 .008 20 .00 0 .08 0 .40 0 .16 0 .40 0 .08 0 .08 0 .24 700 .00 350 .00 350 .00 28 .68 600 .00 600 .00 650 .00 2,500 .00 50 .00 75 .00 625 .00 50 .00 150 .00 350 .00 550 .00 175 .00 100 .00 62 .14 100 .00 700 .00 350 .00 350 .00 28 .68 600 .00 600 .00 650 .00 2,500 .00 50 .00 75 .00 625 .00 50 .00 150 .00 350 .00 550 .00 175 .00 100 .00 62 .14 100 .00 700 .00 350 .00 350 .00 22 .00 600 .00 600 .00 650 .00 2,500 .00 50 .00 75 .00 625 .00 150 .00 350 .00 550 .00 175 .00 100 .00 50 .00 100 .00 1,400 .00 700 .00 700 .00 57 .36 1200 .00 1,200 .00 1,300 .00 5,000 .00 100 .00 150 .00 1,250 .00 100 .00 300 .00 700 .00 1,100 .00 350 .00 200 .00 100 .00 200 .00 800 .00 1,300 .00 1,300 .00 119 .14 1,500 .00 1,000 .00 200 .00 200 .00 800 .00 750 .00 250 .00 700 .00 1,000 .00 1,000 .00 500 .00 200 .00 200 .00 100 .00 360 .40 500 .00 400 .00 400 .00 60 .00 100 .00 800 .00 1,800 .00 250 .00 400 .00 150 .00 150 .00 1,650 .00 800 .00 150 .00 1,700 .00 250 .00 350 .00 100 .00 500 .00 300 .00 11300 Graces* powder, mg/L 11605 Graces* supplemented 1X liquid, mg/L 11595 Graces* unsupplemented 1X liquid, mg/L 11667 Graces* 2X liquid, mg/L 11405 IPL-41L 1X liquid, mg/L 21720 Schneiders 1X liquid
* Grace, T .D . (1962) Nature 195, 788 . Schneider, Imogene (1964) Exp. Zool. 156:1, 91 . Revision of original formulation, Schneider, Imogene (1965) personal communications . Original formulations list Na2HPO 4 at 700 .00 mg/L . Revised formulation believed by author to possess better buffering capacity and better nutritive properties . ** Increased from 2,240 .00 mg/L upon recommendation of Dr . Vaughn .
102
Insect Medium Supplements

4% Agarose Gel
Recommended storage conditions: 2C to 8C .
Cellfectin Reagent
Cellfectin II Reagent is a proprietary, cationic lipid formulation in membrane filtered water suitable for DNA transfection into insect cells . Cellfectin II Reagent is an improved version of the Cellfectin Reagent and delivers the same performance as Cellfectin with a faster protocol . Transfection using Cellfectin II results in consistent and efficient transfection of Sf9, Sf21, and High Five cells in Bac-to Bac, BaculoDirect, and InsectSelect Expression Systems from Invitrogen or equivalent systems . Cellfectin Reagent can also be used to transfect adherent or suspension mammalian cells . Transfection protocols for Sf9 and Sf21 cells are described in this manual . For transfecting High Five insect cells and mamamalian cells with Cellfectin II, refer to the transfection selection tool at www .invitrogen .com/transfection Important Guidelines for Transfection Cellfectin II Reagent is a lipid suspension that may settle with time . Mix thoroughly by inverting the tube 5 10 times before use . The DNA:lipid complex formation time is shorter (~15-30 minutes) when using Cellfectin as compared to Cellfectin Reagent Perform transfection using the appropriate protocol for the system and cell type that you are using to obtain the best results . Do not add antibiotics to media during transfection as this will cause cell death . Transfection complexes must be formed in serum-free medium Test seum-free medium for compatibility with Cellfectin II since some serum-free formulations may inhibit lipid-mediation transfection Optimizing Transfections To optimize the highest transfection efficiency and minimize non-specific effects, optimize transfection conditions by varying cell density, DNA and Cellfectin concentrations, and transfection incubation time .
Chemically Defined Lipid Concentrate

Chemically Defined Lipid Concentrate is a concentrated lipid emulsion designed to reduce or replace fetal bovine serum in cell culture media for a wide variety of applications, including growth and maintenance of CHO, hybridoma, and insect cells in culture; monoclonal antibody production by hybridomas; and viral expression in insect cells . The components of this medium supplement are chemically defined . For product formulation please refer to the tables on page 162 . Insect cell culture tested . Recommended storage conditions: 2C to 8C, in the dark . Attention: This material is hazardous . All persons using this product should review the Material Safety Data Sheet before handling .
Pluronic F-68, 10% (100X)

Prepared in distilled water . Insect cell culture tested . Recommended storage conditions: 15C to 30C .
Quantity 1 ml 125 l Cat. no. 24040-032 Cat. no. 10362-100 10362-125
Quantity 100 ml
Yeastolate Ultrafiltrate (50X), liquid

Yeastolate Ultrafiltrate is formulated specifically for use with IPL-41 Insect Medium to support the growth of Spodoptera frugiperda (Sf9) . Prepared with 200 g/L of yeastolate in distilled water . Insect cell culture tested . Performance testing: Performance tested in a growth assay using Sf9 cells .
103
Transfecting Sf9 or Sf21 with Baculovirus

Use the following procedure to transfect Sf9 or Sf21 insect cells cultured in Graces Insect Medium, supplemented, containing 10% FBS in a 6-well format . All amounts and volumes are given on a per well basis . 1 . If the cell density is in the range o 1 .5-2 .5 x 106 cells ml with 95% viability, and the medium is without antibiotics, proceed to step 1a . If the cell density is not in this range or the medium contains antibiotics, follow steps 1b-1c . a . Add 2ml of Graces Insect Medium, Unsupplemented (without antibiotics and serum) in each well . Plate 8 x 105 Sf9 or Sf21 cells per well . Do not change medium or wash the cells . The medium carried over to the next step will enhance the transfection efficiency . Allow cells to attach for 15 minutes at room temperature in the hood . Proceed to step 2 . b . Prepare 10ml plating medum by mixing 1 .5ml Graces Insect Medium, Supplemented, containing 10%FBS (without antibiotics) and 8 .5ml Graces Insect Medium, Unsupplemented (without FBS and antibiotics) . c . Plate 8 x 105 Sf9 or Sf21 cells per well . Allow cells to attach for 15mins at room temperature . DNA in the hood . Remove the medium . Add 2 .5ml plating medium from step 1b per well . Proceed to step 2 . 2 . For each transfection sample, prepare complexes as follows: Bac-to bac System a . Mix Cellfectin before use and dilute 8 l in 100 ul Graces Insect Medium, Unsupplemented . Vortex briefly to mix Note: This mixture can be kept at room temperature for up to 30 minutes . b . Dilute 1ug Graces Insect Medium, Unsupplemented . Mix Gently .
c . Combine the diluted DNA with diluted Cellfectin II (total volume is ~ 210l) . Mix gently and incubate for 15 30 minutes at room temperature .
Baculo Direct System

a . Prepare Transfection Mix A: Cellfectin II 8l Graces Insect Medium 100ul Unsupplemented Note: This mixture can be kept at room temperature for up to 30 minutes . b . Prepare Transfection Mix B: LR recombination reaction 10l Graces Insect Medium 100l Unsupplemented c . Combine Transfection mixtures A and B . Mix gently and incubate for 25 35 minutes at room temperature . 3 . Add-210ul DNA mixture or transfection mixture (Step 2c) dropwise onto the cells from Step 1a or 1c . Incubate cells at 27oC for 3-5 hours . 4 . Remove the transfection mixture and replace with 2ml of complete growth medium(e .g . Graces Insect Medium, Supplemented and 10% FBS) . Use of antibiotics is optional for bac-to bac system . Use complete medium with 100uM ganciclovir for Baculodirect system . 5 . Incubate cells at 27 0C for 72 hours or until you see signs of viral infection .
104
Transfecting Sf9 or Sf21 with Plasmid DNA

Use this procedure to transfect Sf9 and Sf21 Insect cells in a 6-well format in Sf900 III SFM, Sf-900, or Graces Insect Medium . All amounts and volumes are given on a per well basis . 1 . For log phase cells cultured in Sf-900 II SFM or Sf-900 III SFM without antibiotics: Plate 8 x 105 Sf9 or Sf21 cells per well in 2ml Sf-900 II SFM or Sf-900 III SFM . Allow cells to attach for 15 minutes at room temperature . Proceed to Step 2 . For cells cultured in of Graces Insect Medium: If the cell density is not in this range or the medium contains antibiotics, follow steps 1b 1c . a . Add 2ml of Graces Insect Medium, Unsupplemented (without antibiotics and serum) in each well . Plate 8 x 105 Sf9 or Sf21 cells per well . Do not change medium or wash the cells . The medium carried over to the next step will enhance the transfection efficiency . Allow cells to attach for 15 minutes at room temperature in the hood . Proceed to Step 2 . b . Prepare 10ml plating medium by mixing 1 .5ml Graces Insect Medium, Supplemented, containing 10% FBS (without antibiotics) and 8 .5ml Graces Insect Medium, Unsupplemented, (without FBS and antibiotics) . c . Plate 8 x 105 Sf9 or Sf21 cells per well . Allow cellsnto attach for 15 minutes at room temperature in the hood . Remove the medium . Add 2 .5ml plating medium from step 1b per well, Proceed to step 2 . 2 . For each transfection sample, prepare complexes as follows: a . Dilute 1 g plasmid DNA in 100 l Graces Insect Medium, Unsupplemented (without antibiotics and serum) . Vortex briefly to mix, Incubate at room temperature for 15-30 minutes . Optional: If you are using SFM, add 5 l PLUS Reagent (Cat .no . 11514-015) to the diluted DNA to improve transfection efficiency . b . Mix Cellfectin II before use, then dilute 8 l Cellfectin II in 100l Graces Insect Medium, unsupplemented (without antibiotics and serum) . Vortex briefly to mix . Incubate at room temperature for 15 30 minutes. c . Combine the diluted DNA diluted Cellfectin II (total volume is ~210 l) . Mix gently and incubate for 15 30 mins .
3 . Add ~ 210 l DNA lipid mixture (Step 2c) dropwise onto the cells from Step 1a or 1c . 4. For Sf-900 II SFM or Sf-900 III SFM: Incubate cells at 27oC for 2448 hours and test for transgene expression . There is no need to replace the medium . 5. For Graces medium: Incubate cells at 27oC for 3-5 hours . Remove the transfection mixture and replace with 2ml of complete growth medium (e .g . Graces, Supplemented, and 10% FBS)> Addition of antibiotics to the growth medium is optional . Incubate cells at 27oC for 2448 hours and test for transgene expression . Quality Control The Certificate of Analysis (CofA) Provides detailed quality control information for each product . Certificates of analysis are available from www .invitrogen .com/support and search for the Certificate of Analysis by product lot number, which is printed on the box Limited Use Label No.326: Cellfectin II Transfection Reagent This product is subject to U .S Patents 4, 897, 355 and 5, 550, 289 . This Product s licensed for use in in vitro delivery of proteins, peptides, or other macromolecules only . For a license to use the product in any other field-of-use, including in vitro uses, contact Roche Palo Alto LLC>< 3431 Hillview Avenue, Palo Alto CA 04304-1397 2008 Invitrogen Corporation . All rights reserved . For research use only . Not intended for any animal or human therapeutic or diagnostic use .


Hybridoma and Monoclonal Antibody Products
CD Hybridoma Medium (1X), liquid
CD Hybridoma Medium is a chemically defined, protein-free medium optimized for the growth of a variety of hybridomas and myelomas and the production of monoclonal antibodies in stationary or agitated suspension systems .1,2 CD Hybridoma Medium contains no proteins of animal, plant, or synthetic origin . There are also no undefined lysates or hydrolysates in the formulation . Formulated without L-glutamine for increased stability; add 40 ml of 200 mM L-glutamine or 40 ml of GlutaMAX-I Supplement per 1,000 ml of medium prior to use . CD Hybridoma Medium is not optimized for lipid-dependent or cholesterol-dependent cultures such as NS0-derived lines . Please inquire about options for those applications . Recommended storage conditions: 2C to 8C, in the dark . Notes: A Hybridoma Medium Master file has been submitted to the FDA . Permission to cross reference the Master file may be obtained by contacting Invitrogen Technical Support or you local Sales Representative .
Related Products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .140 250X Cholesterol Lipid Concentrate for growing NS0 and other lipid dependent cell lines . . . . . . . . . . . . . . 30 Hybridoma reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .159 Distilled Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .152
Hybridoma-SFM (1X), liquid

Hybridoma-SFM is a very lowprotein (containing 20 g/ml protein as insulin and transferrin) medium optimized for the growth of a variety of hybridomas and the production of monoclonal antibodies . Most hybridomas require only minor sequential adaptation or no adaptation to this medium . The very low protein content facilitates monoclonal antibody purification . Recommended storage conditions: 2C to 8C, in the dark . Notes: A Hybridoma Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
References
1 . Gorfien, S .F ., Godwin, G .P ., Samrock, R ., and Tilkins, M .L . (1999) FOCUS 21, 30 . 2 . Csirke, B ., Godwin, G .P ., and Gorfien, S .F . (1999) FOCUS 21, 33 .
Reference
1 . Mullins, T .D ., Dzimian, J .L ., Fike, R .M ., and Weiss, S .A . (1991) FOCUS 13, 91 .
Quantity 1,000 ml
Contains no L-glutamine or phenol red .
Quantity Cat. no. 11279-023 500 ml 1,000 ml
Cat. no. 12045-084 12045-076
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 250X Cholesterol Lipid Concentrate for growing NS0 and other lipid dependent cell lines . . . . . . . . . . . . .106 Hybridoma reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .166
Related products Page Hybridoma reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .166
250X Cholesterol Lipid Concentrate

250X Cholesterol Lipid Concentrate is a chemically defined, protein-free supplement used for growing NS0 and other lipid-dependent cell lines to high densities while supporting growth and protein expression without complicating downstream purification . The supplement can be added to 1X media such as CD Hybridoma and filtered with minimal loss of lipids . Custom formulations and packaging configurations as well as supplements used in feeding strategies are available through our Custom Products Services group . Recommended storage conditions: 2C to 8C, in the dark .
CD Hybridoma AGT
CD Hybridoma AGT is a chemically defined, protein-free medium in a new easy-to-use granular format . CD Hybridoma AGT is optimized for the growth of a variety of hybridoma and myelomas and the production of monoclonal antibodies in stationary or agitated suspension systems . Recommended storage conditions: 2C to 8C, in the dark .
Reference
1 . Gorfien, S ., Paul, W ., Walowitz, J ., Keem, R ., Biddle, W . and Jayme, D . (2000), Biotechnol. Prog. 16, 682-687 .
106
PFHM-II Protein-free Hybridoma Medium (1X), liquid
HAT Supplement (50X), liquid

HAT Supplement (50X) is a liquid mixture of sodium hypoxanthine, aminopterin, and thymidine . After dilution, HAT is suitable for use as a post-fusion selective medium to eliminate unfused or selffused HGPRT-myeloma cells . Hypoxanthine and thymidine supply preformed purines and pyrimidines for DNA synthesis by hybridomas via the salvage pathway that utilizes HGPRT contributed by the fused spleen cell . Aminopterin, a folic acid antagonist, inhibits the de novo nucleoside biosynthesis pathway . Concentration at 50X: 5 mM sodium hypoxanthine, 20 M aminopterin, 0 .8 mM thymidine . Recommended storage conditions: 5C to 20C .
PFHM-II is a protein-free, ready-to-use medium that contains no polypeptide growth or attachment factors, or mediators that may complicate downstream processing and final product purification .1 PFHM-II performs as well as serum-supplemented media for MAb production and also may be used as a growth medium . In instances where antibiotics are required, most general antibiotics are compatible with PFHM-II, including penicillin/streptomycin, gentamicin, anti-PPLO, linocin, and Fungizone Reagent . Do not use kanamycin sulfates or neomycin sulfates . Recommended storage conditions: 2C to 8C, in the dark .
Fungizone is a trademark of E .R . Squibb & Sons, L .L .C .
Reference
1 . Fike, R ., Pfohl, J ., Jayme, D ., and Weiss, S .A . (1990) FOCUS 12, 79 .
Quantity 1,000 ml
Cat. no. 12040-077
HT Supplement (100X), liquid

HT Supplement (100X) liquid is a mixture of sodium hypoxanthine and thymidine . After dilution, HT Supplement is suitable for use as a rescue medium . The supplement provides preformed purines and pyrimidines to overcome the effects of residual intracellular aminopterin . Once the de novo biosynthesis pathway for nucleosides has been re-established, HT supplementation can be discontinued . Concentration at 100X: 10 mM sodium hypoxanthine, 1 .6 mM thymidine . Recommended storage conditions: 2C to 8C .
Medium NCTC-109 (1X), liquid

NCTC-109 (1X) is a synthetic medium that is suitable for general use in generating and maintaining hybridoma cells . This medium typically requires the addition of a growth factor cocktail or serum . Recommended storage conditions: 2C to 8C . Intended use: In vitro diagnostic (IVD) .
Quantity 500 ml
Contains no L-glutamine .
Quantity Cat. no. 21340-054 50 ml Cat. no. 11067-030
Customized Hybridoma Medium Products

Hybridoma and Monoclonal Antibody products, along with DMEM/F-12, IMDM/F-12, and your in-house formulations are available on a make-toorder basis . We realize that not all needs are alike, so weve developed the capabilities to provide the desired configuration (formulation and packaging) of product type, as well as the operational systems that can provide these products in a timely fashion . Please contact our Custom Products Services group for assistance in selecting the products and packaging that best meet your requirements . Additional considerations for optimizing hybridoma systems include the use of nutrient supplements (specially developed for your system or general) and glutamine dipeptides (see GlutaMAX Supplement, page 54) for enhanced stability .
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Protein A Agarose
Protein A Agarose is suitable for purifying IgG molecules from a variety of species . Protein A Agarose also can be used for fractionation of IgG fragments, fractionation of cells tagged with IgG, and isolation of antigenantibody complexes . In addition, Protein A interacts with some classes of IgM and IgA . The matrix consists of purified Protein A covalently attached to cross-linked 4% agarose beads using the cyanogen bromide method . Protein A Agarose can bind >18 mg of human IgG per ml of gel . Performance and quality testing: Binding capacity for human IgG is determined by affinity chromatography . Concentration in storage buffer: 45% to 55% suspension in 20 mM sodium phosphate (pH 7 .2), 0 .15 M NaCl, 0 .02% (w/v) merthiolate . Recommended storage conditions: 4C .
Protein A Agarose is manufactured under license from Amersham Pharmacia Biotech, Inc .
Streptavidin Agarose
Streptavidin Agarose is suitable for isolating biotinylated molecules and complexes with biotinylated components, including antibodies, antibody-antigen complexes, ligands and receptors, nucleic acids, and nucleic acid-protein complexes . It is especially useful as an immunoprecipitation reagent for antibodies that do not bind well to Protein A . The matrix consists of streptavidin covalently attached to 4% agarose beads through a 15-atom spacer arm . Streptavidin Agarose can bind 3 to 8 mg of biotinylated IgG per ml of gel . Streptavidin Agarose is supplied as 10 ml of a 50% slurry containing 5 ml of packed gel bed in 0 .01 M sodium phosphate (pH 7 .2), 0 .15 M NaCl, and 0 .02% sodium azide . Recommended storage conditions: 4C .
Quantity 5 ml Cat. no. SA100-04
Recombinant Protein G (rProtein G) Agarose

Recombinant Protein G (rProtein G) Agarose is suitable for purification of IgG antibodies using column chromatography or immunoprecipitation methods . This matrix, which contains a genetically engineered form of streptococcal Protein G, is designed to eliminate binding of serum albumin while maintaining efficient binding of the Fc region of IgG isotypes . rProtein G is produced in E. coli and, after purification, is covalently immobilized on 4% agarose beads by cyanogen bromide . rProtein G Agarose binds to a broader range of IgG isotypes than react with Protein A . rProtein G Agarose can bind >18 mg of human IgG per ml of gel . The binding characteristics and stability of rProtein G Agarose facilitate rapid and repeated use for affinity chromatography . rProtein G Agarose is supplied as a 50% suspension in PBS (pH 7 .0), 20% ethanol . Recommended storage conditions: 4C .
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Cellular Studies
Bronchial Epithelial Products
LHC Medium (1X), liquid
Formulated for the growth of bronchial epithelial cells with further supplements . LHC serum-free media are the most widely used products for culture of bronchial epithelial cells . For maximum flexibility and convenience, we offer these products with or without supplementation . Recommended storage conditions: 2C to 8C .
Recommended applications
Complete media (for cell culture for the following research/disease areas: asthma, allergies, lung cancer, cystic fibrosis, pulmonary and esophageal cancer*) Media supplements Typical cells cultured Retinoic acid IB3-1, S9, C38 BEAS-2B, BZR, BBM, Het-1A, NHBE BBE Basal media (for flexible customization) Epinephrine Gentamicin Other supplements LHC product LHC-8 LHC-8 w/o gentamicin LHC-9 LHC Basal
* For research use only . Includes insulin, hydrocortisone, EGF, transferrin, bovine pituitary extract, and T3 .
Product LHC Basal Medium (1X), liquid LHC-8 Medium (1X), liquid Quantity 500 ml 1,000 ml 500 ml Cat. no. 12677-019 12677-027 12678-017
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Dermatology Products
Defined Keratinocyte-SFM (1X), liquid
Defined Keratinocyte-SFM is a serum-free medium optimized for the isolation and expansion of human keratinocytes without the need for bovine pituitary extract (BPE) supplementation or the use of fibroblast feeder layers .1,2 The growth-promoting activity of BPE has been replaced by the inclusion of growth promoting agents which enhance consistent product performance . These growth-promoting agents also maintain cell morphology, physiological markers, and growth . The medium is formulated to inhibit fibroblast proliferation and has a low calcium concentration (<0 .1 mM) allowing for the isolation and culture of highly enriched populations of undifferentiated keratinocytes . Recommended storage conditions: medium, 2C to 8C, in the dark; supplement, 5C to 20C . Shelf life after supplementation: 10 weeks . Notes: This formulation contains some human and animal-origin components . A Defined Keratinocyte-SFM Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen nical Support or your local Sales Representative . Defined Keratinocyte-SFM consists of a basal liquid medium and frozen supplement which is added at the time of use . The fully supplemented medium has a shelf life of 10 weeks under normal use conditions . Product performance is confirmed by evaluation of human primary keratinocyte growth .
Keratinocyte-SFM (1X), liquid

Keratinocyte-SFM is a serum-free medium (with BPE) optimized for the cultivation of primary and passaged human keratinocytes . 1,2 Keratinocyte-SFM can be used with or without an irradiated fibroblast feeder layer . The formulation retards fibroblast overgrowth, thereby allowing the study of highly pure populations of keratinocytes . The low calcium ion concentration (<0 .1 mM) permits greater cell growth and slows cell differentiation . Keratinocyte-SFM is supplied with prequalified human recombinant Epidermal Growth Factor 1-53 (EGF 1-53), and Bovine Pituitary Extract (BPE) . Separate packaging of these components allows flexibility in determining the supplement concentrations most suited to the application . Recommended storage conditions: medium, 2C to 8C, in the dark; supplements: 5C to 20C .
Quantity 500 ml
Cat. no. 17005-042
References
1 . Daley, J .P ., Epstein, D .A ., and Hawley-Nelson, P . (1990) FOCUS 12, 68 . 2 . Donovan, J . (1998) FOCUS 20, 38 .
Keratinocyte-SFM (1X), liquid (without CaCl2)

Keratinocyte-SFM is supplied with prequalified human recombinant Epidermal Growth Factor 1-53 (EGF 1-53), and Bovine Pituitary Extract (BPE) in separate packaging . Components are not sold separately . Recommended storage conditions: medium, 2C to 8C, in the dark; supplements: 5C to 20C . Shelf life after supplementation: 2 weeks .
References
1 . Judd, D .A ., Battista, P .J . and Behm, D .D . (1997) FOCUS 19, 2 . 2 . Donovan, J . (1998) FOCUS 20, 38 .
Quantity 500 ml
Contains L-glutamine but no calcium chloride .
Cat. no. 37010-022
Related products Page Cell Dissociation Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .158 Primary Human Keratinocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .179
Keratinocyte Starter Kit

The Keratinocyte Starter Kit includes Defined Keratinocyte-SFM Medium, growth supplement, and Primary Human Keratinocytes . Recommended storage conditions: medium, 2C to 8C, in the dark; supplement, 5C to 20C; cells, 125C to 200C (liquid nitrogen) .
Hematology-Oncology Research Products (AIM V Media)

Serum-free media developed for research applications using various differentiated cell types may also be evaluated for biomedical research applications . Although products and components are screened for absence of contaminants, we recommend that users exercise caution in handling all products containing materials of human/animal origin and encourage handling practices consistent with materials that are potentially infectious . It is the exclusive responsibility of the end user to qualify/validate these products for specific applications .
Quantity 1 kit Cat. no. 12568-010
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AIM V Medium for Lymphocyte Culture

AIM V Medium (serum-free lymphocyte medium) was designed specifically for research purposes in adoptive immunotherapy, a method of cancer treatment explored in clinical trials . The medium has, however, been found to be useful in a growing list of applications, including culturing functionally differentiated lymphoid cell lines, investigating signal transduction pathways, and performing applications requiring component definition . The medium has applications in the following areas: lymphokine-activated killer (LAK) cell induction and function,17 cytokine and antibody activity in LAK cell induction,812 long-term cultivation of functional human macrophages,13 immunotherapy of murine hepatic metastases,14 lymphocyte receptor studies,15 and ADCC-dependent lymphocyte activation .16 As shown in the table, AIM V Medium has been used for serum-free growth of many cell types . The medium may also have applications in the following areas of research: NK cells and NK-like effector populations; T cell activation; cytokine regulation of effector cells; cytokine interactions and cascades; cytokine receptor and postreceptor events in lymphoid effector cells; macrophage activation; effector cells in antitumor immunity; mechanisms of cell-mediated cytotoxicity; in vitro effects of lymphokines, antibodies, and immunotherapy; hybridoma growth and monoclonal antibody production; AIDS research; production of HIV proteins (e .g ., gP160 and gP120); and definition and isolation of regulatory proteins produced by activated monocytes and macrophages .
References
1 . Muul, L .M ., Nason-Burchenal, K ., Hyatt, C ., Schwarz, S ., Slavin, D ., Director, E .P ., and Rosenberg, S .A . (1987) J. Immunol. Methods 105, 183 . 2 . Jadus, M .R ., Thurman, G .B ., Mrowca-Bastin, A ., and Yanelli, J . (1988) J. Immunol. Methods 109, 169 . 3 . Grimm, E .A ., Owen-Schaub, L .B ., Loudon, W .G ., and Yagita, M . (1988) Ann. N.Y. Acad. Sci. 532, 788 . 4 . Beckner, S .K ., Maluish, A .E ., and Longo, D .L . (1987) Cancer Res. 47, 5504 . 5 . Ryan, T ., Atkins, M .B ., Mier, J .W ., and Beckman, E .M . (1989) Transfusion 29, 491 . 6 . Knazek, R .A ., Ulu, Y-W ., Aebersold, P .M ., and Rosenberg, S .A . (1990) J. Immunol. Methods 127, 29 . 7 . Haas, G .P ., Solomon, D ., and Rosenberg, S .A . (1990) Cancer Immunol. Immunother. 30, 342 . 8 . Owen-Schaub, L .B ., Gutterman, J .V ., and Grimm, E .A . (1988) Cancer Immunol. Immunother. 29, 193 . 9 . Yang, S .C ., Owen-Schaub, L .B ., Grimm, E .A ., and Roth, J .A . (1989) Cancer Immunol. Immunother. 29, 193 . 10 . Grimm, E .A ., Crump III, W .L ., Durett, A ., Hester, J .P ., Lagoo-Deenadalayan, S ., and Owen-Schaub, L .B . (1988) Cancer Immunol. Immunother. 27, 53 . 11 . Crump III, W .L ., Owen-Schaub, L .B ., and Grimm, E .A . (1989) Cancer Res. 49, 149 . 12 . Yang, S .C ., Owen-Schaub, L .B ., Grimm, E .A ., and Roth, J .A . (1988) Surg. Forum 39, 454 . 13 . Helinski, E .M ., Bielat, K .L ., Ovak, G .M ., and Pauly, J .L . (1988) J. Leukocyte Biol. 44, 111 . 14 . Lafreniere, R ., Borkenhagen, K ., Bryant, L .D ., and Huchcroft, S . (1989) Cancer Res. 49, 2409 . 15 . Owen-Schaub, L .B ., Crump III, W .L ., Morin, G .I ., and Grimm, E .A . (1989) J. Immunol. 143, 2336 . 16 . Masacci, G ., Weroall, P ., Nielsen, J ., Kierulff-Nielsen, H ., Wizzell, H ., and Mellstedt, M . (1989) Hybridoma 8, 507 .
Cell Types Successfully Cultivated Using Serum-free AIM V Medium

Cell line/type
Human primary lymphoid cells T cells T cells T cells CD8+ T lymphocytes Macrophages Human monocytes Transformed lymphoid lines HUT78 HL-60 COS cells (monkey) YAC-1 (mouse) Murine lymphoma Myeloid cells Hybridoma Leukemic cells Adherent cells Oral epithelial Human fibroblasts VERO Growth Growth Growth DMEM RPMI 1640 MEM Retrovirus research Nuclear transcription research Protein secretion IL-2 activation Growth Growth Monoclonal antibody production Immune regulation RPMI 1640, IMDM DMEM DMEM RPMI 1640 DMEM RPMI 1640 RPMI 1640, IMDM RPMI 1640 Activation Cloning Retrovirus research CTL induction Differentiation Growth RPMI 1640 RPMI 1640 RPMI 1640 RPMI 1640 RPMI 1640, IMDM RPMI 1640
Application
Media used previously*
* Supplemented with 10% fetal bovine serum unless otherwise indicated . Supplemented with 10% human serum . Supplemented with 20% newborn calf serum . Supplemented with 10% newborn calf serum .
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AIM V Medium, liquid (research grade)

Performance testing: Performance tested in a growth assay using Jurkat cells (ATCC Jurkat T1B-152) . Viral testing: Raw components of human origin are nonreactive, at the donor level, for Hepatitis B surface antigen (HBsAG), antibody to Hepatitis C virus (anti-HCV), antibodies to Human Immunodeficiency Viruses 1 and 2 (anti-HIV 1 and 2), and syphilis, as determined by currently approved testing procedures . Raw material components of human origin are also tested for alanine aminotransferase (ALT) enzyme, and found to be within normal limits . Where possible, such components are further processed by heat treatment in liquid form at 60C for 10 hours by validated methods demonstrated to inactivate adventitious viral contaminants (performed by our raw material vendors) . Recommended storage conditions: 2C to 8C, in the dark .
AIM V Medium, liquid (therapeutic grade)

Performance testing: Performance tested in a growth assay using Jurkat cells (ATCC Jurkat T1B-152) . Viral testing: Raw material components of human origin are nonreactive, at the donor level, for Hepatitis B surface antigen (HBsAG), antibody to Hepatitis C virus (anti-HCV), antibodies to Human Immunodeficiency Viruses 1 and 2 (anti-HIV 1 and 2), and syphilis, as determined by currently approved testing procedures . Raw material components of human origin are also tested for alanine aminotransferase (ALT) enzyme, and found to be within normal limits . Where possible, such components are further processed by heat treatment in liquid form at 60C for 10 hours by validated methods demonstrated to inactivate adventitious viral contaminants (performed by our raw material vendors) . Intended uses: For human ex vivo tissue and cell culture processing applications . Recommended storage conditions: 2C to 8C, in the dark . Intended uses: For human ex vivo tissue and cell culture processing applications . Notes: An AIM V Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
Product AIM V Medium, liquid (research grade, with HSA) Quantity 500 ml 1,000 ml Cat. no. 12055-091 12055-083
Contains L-glutamine, streptomycin sulfate at 50 g/ml, and gentamicin sulfate at 10 g/ml .
Product AIM V Medium, liquid (research grade, with BSA)
Quantity 500 ml
Cat. no. 31035-025
Quantity 1,000 ml 10 L Cat. no. 087-0112DK 087-0112BK
Contains AlbuMAX supplement (bovine serum albumin) substituted for human serum albumin, L-glutamine, streptomycin sulfate at 50 g/ml, and gentamicin sulfate at 10 g/ml .
Contains L-glutamine, 50 g/ml streptomycin sulfate, and 10 g/ml gentamicin sulfate .
112
Hematopoiesis Products
StemPro-34 SFM (1X), liquid
S tem Pro -34 SFM is a serum-free medium specifically formulated to support the development of human hematopoietic cells in culture .1 StemPro-34 SFM can be used to study hematopoietic cells from bone marrow, peripheral blood, or neonatal cord blood . StemPro-34 SFM is performance-tested on normal human bone marrow . It is manufactured with components of human, recombinant, or synthetic origin . StemPro-34 SFM consists of a basal liquid medium and frozen supplement (StemPro-Nutrient Supplement) which is added at the time of use . Recommended storage conditions: medium, 2C to 8C; supplement, 5C to 20C . Notes: An intended use disclosure is required for purchase of StemPro-34 SFM . For further regulatory information on the uses of this product, please contact the Tech-LineSM .
Hepatocyte Products
Gibco hepatocyte products include a complete range of serum-free media and reagents designed for the isolation and maintenance of hepatocytes, while sustaining the normal phenotypic expression including xenobiotic metabolism .1
Reference
1 . Alfonso, J .L ., Samrock, R ., and Price, P . (1995) FOCUS 17, 6 .
Hepatozyme-SFM (1X), liquid

Hepatozyme-SFM is a serum-free medium for the long-term maintenance of hepatocyte phenotypic expression, including active and inducible forms of cytochromes P450 and active phase II enzymes . Recommended storage conditions: 2C to 8C, in the dark .
Quantity 500 ml
Contains no phenol red .
References
1 . Daley, J .P ., Dadey, B .M ., Wysocki, M .G ., Caligiuri, M .A ., and Biddle, W .C . (1996) FOCUS 18, 62 . 2 . Taylor, M . et al . (2001) Blood 98, 1195-1199 . 3 . Szilvassy, S . et al . (1999) Blood 93, 1557-1566 . 4 . Delehanty, L . et al . (2003) Blood 101, 1744-1751 . 5 . Kirshenbaum, A ., et al . (1999) Blood 94, 2333-2342 . 6 . Szilvassy, S ., Meyerrose, T and Grimes, B . (2000) Blood 95, 2829-2837 . 7 . Yahata, T ., et al . (2003) Blood 101, 2905-2913 . 8 . Wang, J ., et al . (2003) Blood 101, 2924-2931 . 9 . Kolbus, A ., et al . (2003) Blood 102, 3136-3146 . 10 . Goyama, S ., et al . (2004) Blood 104, 3558-3564 .
Cat. no. 17705-021
Liver Digest Medium (1X), liquid

Liver Digest Medium is a collagenase-dispase medium for dissociation of viable liver cells . Recommended storage conditions: 5C to 20C, in the dark .
Liver Perfusion Medium (1X), liquid

Liver Perfusion Medium is a buffered, balanced salt solution formulated to cleanse the liver of blood, prevent clotting, and initiate loosening of cell-to-cell contact . Recommended storage conditions: 2C to 8C, in the dark .
Macrophage-SFM (1X), liquid

Macrophage-SFM is a serum-free medium designed for the culture of human peripheral blood monocytes and macrophages .1 Recommended storage conditions: 2C to 8C, in the dark .
Reference
1 . Epstein, D ., Wysocki, M ., Biddle, W ., Streck, R .J ., Rolf, M .M ., and Pauly, J .L . (1991) FOCUS 13, 120 .
Quantity 500 ml
Contains L-glutamine, but no antibiotics .
Hepatocyte Wash Medium

Cat. no. 12065-074
Hepatocyte Wash Medium is an enriched Williams Medium E for the reduction of cell membrane leakage, ease of centrifugation, and high viability . Recommended storage conditions: 2C to 8C, in the dark .
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Human Mammary Epithelial Products
HuMEC Medium
Optimized specifically for culture of human mammary epithelial cells, HuMEC Medium is a serum-free formulation with frozen supplements containing growth factors and other components . This translates to greater experimental validity, more reproducible results, and reduction of lot-to-lot variability and aberrations that can occur with the addition of serum . Human mammary epithelial cells cultured in HuMEC Medium have gene expression profiles consistent with normal mammary epithelial cells, as tested on a cDNA microarray . HuMEC Ready Medium includes HuMEC Basal Serum-free Medium and a HuMEC Supplement Kit . The HuMEC Supplement Kit includes 5 ml of a supplement mix containing epidermal growth factor, hydrocortisone, isoproterenol, transferrin, and insulin, and 25 mg of bovine pituitary extract . Applications: HuMEC Medium has been performance tested in a growth assay using normal human mammary epithelial cells . HuMEC Medium allows the culture of human mammary epithelial cells under conditions optimized for breast cancer research, providing researchers in drug discovery and other fields greater insight into carcinogenic progression . Recommended storage conditions: basal medium, 2C to 8C; supplement mix, 5C to 20C .
Neurobiology Products
Neurobasal Media
Neurobasal Media are basal media formulated to specifically meet neuronal cells special requirements . They allow for long-term maintenance of the normal phenotype and growth of neuronal cells, and maintain pure populations of neuronal cells without the need for an astrocyte feeder layer . Neurobasal Medium is for long-term growth of fetal hippocampal neurons and many other neurons of the CNS .1 Neurobasal-A Medium is for long-term growth and viability of rat postnatal and adult hippocampal and cortical neurons of the CNS .2 Prior to use, Neurobasal Media must be supplemented with either a serum-free supplement (such as B-27 or N-2) or serum, and 0 .5 mM L-glutamine or GlutaMAX-I Supplement . For initial plating, add 25 M glutamic acid . Recommended storage conditions: 2C to 8C, in the dark .
References
1 . Brewer, G .J ., Torricelli, J ., Evege, E .K ., and Price, P . (1994) FOCUS 16, 6 . 2 . Brewer, G .J . (1997) J. Neurosci. Methods 71, 145 . 3 . Brewer, G ., Price, P . (2001) Protocols for Neural Cell Culture, 3rd Ed ., Ed .: S . Federoff and A . Richardson, Humana Press, 255264 . 4 . Hermann, A . et al . (2004) J Cell Science 117, 44114422 .
Product HuMEC Basal Serum-free Medium (1X), liquid HuMEC Ready Medium (1X), liquid HuMEC Supplement Kit Quantity 500 ml 500 ml 1 kit Cat. no. 12753-018 12752-010 12755-013
Product Neurobasal Medium (1X), liquid Neurobasal Medium without phenol red (1X), liquid Neurobasal-A Medium (1X), liquid Neurobasal-A Medium without phenol red (1X), liquid Quantity 500 ml 500 ml 500 ml 500 ml Cat. no. 21103-049 12348-017 10888-022 12349-015
Contains no L-glutamine, L-glutamic acid, or aspartic acid .
Related products Page L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .123 Cell Dissociation Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .158 TrypLE Cell Dissociation Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .159 Lipofectamine 2000 Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . 132
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B-27 Serum-free Supplements

B-27 Supplements are 50X liquid, serum-free supplements for growth and long-term viability of hippocampal neurons .1,2 These supplements are used in combination with a basal medium such as Neurobasal Medium .2 B-27 Supplement Minus AO is the complete B-27 Supplement without 5 antioxidants (vitamin E, vitamin E acetate, superoxide dismutase, catalase, and glutathione) that would interfere with studying free-radical damage to neurons . B-27 Supplement Minus Vitamin A is the complete B-27 Supplement without Vitamin A . The deletion of vitamin A in this supplement prevents neural differentiation of stem cells . Recommended storage conditions: 5C to 20C .
Brain-Derived Neurotrophic Factor (BDNF), Human, Recombinant

Brain-Derived Neurotrophic Factor (BDNF), Human, Recombinant is prepared from E. coli, and is supplied lyophilized from distilled water with no additives . Applications: Maintenance of motor neurons and the differentiation of both cholinergic and dopaminergic neurons . Performance and quality testing: Determined to be >96% pure by SDS-PAGE . Functional performance is verified by dose-dependent induction of choline acetyl transferase activity in primary septal cell cultures . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in glass or in dilute solution . Membrane-filtered through 0 .22 m filter .
References
1 . Brewer, G .J ., Torricelli, J .R ., Evege, E .K ., and Price, P .J . (1993) J. Neurosci. Res. 35, 567 . 2 . Brewer, G .J ., Torricelli, J .R ., Evege, E .K ., and Price, P .J . (1994) FOCUS 16, 6 . 3 . Brewer, G ., Price, P . (2001) Protocols for Neural Cell Culture, 3rd Ed ., Ed .: S . Federoff and A . Richardson, Humana Press, 255264 . 4 . Hermann, A . et al . (2004) J Cell Science 117, 44114422 . 5 . Eide, L . and McMurray, C . (2005) BioTechniques 38, 99104 .
Quantity 5 g Cat. no. 10908-010
Product B-27 Serum-free Supplements (50X), liquid B-27 Supplement Minus AO (50X), liquid B-27 Supplement Minus Vitamin A (50X), liquid Quantity 10 ml 10 ml 10 ml Cat. no. 17504-044 10889-038 12587-010
G-5 Supplement (100X), liquid

G-5 Supplement is for the growth and expression of glial cells (normal and tumor) of astrocytic phenotype . For product formulation please refer to the tables on page 162 . Recommended storage conditions: 5C to 20C .
N-2 Supplement (100X), liquid

N-2 Supplement is for the growth and expression of post-mitotic neurons and tumor cells of neuronal phenotype . For product formulation please refer to the tables on page 162 . Recommended storage conditions: 5C to 20C .
115
Nerve Growth Factor 2.5S (NGF 2.5S), Murine, Natural

NGF 2 .5S, Murine, Natural a ~26kDa subunit of the 7S NGF complex, is the 2 .5S subunit responsible for most of the biological activities of NGF . The 2 .5S subunit promotes nerve growth and maintenance of adrenergic neurons in vitro . This preparation of NGF 2 .5S is purified from the submaxillary glands of male mice and is supplied at a concentration of 100 g/ml in 10 mM sodium acetate (pH 5 .0) . Applications: Induction of neurite outgrowth; maintenance of neural cell viability in culture; affects cellular morphology and differentiation; stimulates protein tyrosine . Performance and quality testing: Determined to be >95% pure by SDS-PAGE . Functionality is verified in a neurite outgrowth assay using PC12 cells . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in dilute solution .
ViraPower Lentiviral Expression System

The ViraPower Lentiviral Expression System readily transduces both dividing and non-dividing cell types (neurons), enabling analysis of long-term stable gene expression . Use this system to broaden your ability to analyze gene expression beyond the limitations of traditional transfection reagents or other retroviral systems . Invitrogen is the only commercial supplier of an inducible and promoterless lentiviral expression system .
Product ViraPower Lentiviral Directional TOPO Expression Kit ViraPower Lentiviral Gateway Expression Kit ViraPower Zeo Lentiviral Gateway Expression Kit ViraPower UbC Lentiviral Gateway Expression Kit Quantity 1 kit 1 kit 1 kit 6 g 1 kit 1 kit Cat. no. K4950-00 K4960-00 K4980-00 K4990-00 K4965-00 K5910-00
ViraPower T-Rex Lentiviral Expression System ViraPower Promoterless Lentiviral Gateway Expression System
Nerve Growth Factor 7S (NGF 7S), Murine, Natural

NGF 7S, Murine, Natural, is a 130140-kDa complex that regulates the development and maintenance of sympathetic and some sensory neurons . Murine natural NGF 7S is active on human, mouse, rat, and chicken cells . The NGF 7S is supplied lyophilized from PBS . Applications: Studies of nerve cell growth and differentiation, and biochemical studies of the basis of the various rapid and delayed biological effects of NGF . It can also be used as a medium supplement to support the growth or survival of neuronal cells in culture . Performance and quality testing: Determined to be >95% pure by SDS-PAGE . Functional performance is verified by measuring neurite outgrowth in chick embryo dorsal root ganglia . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in dilute solution . Membrane-filtered through a 0 .22 m filter .
For more information on the ViraPower Lentiviral Expression System, please visit our website at www .invitrogen .com .
116
Stem Cell Research Products

B-27 Supplement Minus AO (50X), liquid
B-27 Supplement Minus AO is the complete B-27 Supplement without 5 antioxidants (vitamin E, vitamin E acetate, superoxide dismutase, catalase, and glutathione) that would interfere with studying free-radical damage to neurons .
Neurobasal-A Medium (1X), liquid

Neurobasal-A Medium is for long-term growth and viability of rat postnatal and adult hippocampal and cortical neurons of the CNS .2 Recommended storage conditions: 2C to 8C, in the dark .
References
1 . Brewer, G .J ., Torricelli, J ., Evege, E .K ., and Price, P . (1994) FOCUS 16, 6 . 2 . Brewer, G .J . (1997) J. Neurosci. Methods 71, 145 . 3 . Brewer, G ., Price, P . (2001) Protocols for Neural Cell Culture, 3rd Ed ., Ed .: S . Federoff and A . Richardson, Humana Press, 255264 . 4 . Hermann, A . et al . (2004) J Cell Sci. 117, 44114422 .
B-27 Supplement Minus Vitamin A (50X), liquid

B-27 Supplement Minus Vitamin A is the complete B-27 Supplement without Vitamin A . The deletion of vitamin A in this supplement prevents neural differentiation of stem cells . Recommended storage conditions: 5C to 20C .
AIM V Media (research grade)

Performance testing: Performance tested in a growth assay using Jurkat cells (ATCC Jurkat T1B-152) . Viral testing: Raw material components of human origin are nonreactive, at the donor level, for Hepatitis B surface antigen (HBsAG), antibody to Hepatitis C virus (anti-HCV), antibodies to Human Immunodeficiency Viruses 1 and 2 (anti-HIV 1 and 2), and syphilis, as determined by currently approved testing procedures . Raw material components of human origin are also tested for alanine aminotransferase (ALT) enzyme, and found to be within normal limits . Where possible, such components are further |processed by heat treatment in liquid form at 60C for 10 hours by validated methods demonstrated to inactivate adventitious viral contaminants (performed by our raw material vendors) .
References
1 . Brewer, G .J ., Torricelli, J .R ., Evege, E .K ., and Price, P .J . (1993) J. Neurosci. Res. 35, 567 . 2 . Brewer, G .J ., Torricelli, J .R ., Evege, E .K ., and Price, P .J . (1994) FOCUS 16, 6 . 3 . Brewer, G ., Price, P . (2001) Protocols for Neural Cell Culture, 3rd Ed ., Ed .: S . Federoff and A . Richardson, Humana Press, 255264 . 4 . Hermann, A . et al . (2004) J Cell Sci. 117, 44114422 .

N-2 Supplement is for the growth and expression of post-mitotic neurons and tumor cells of neuronal phenotype . For product formulation please refer to the tables on page 162 . Recommended storage conditions: 5C to 20C .
AIM V Medium, liquid (research grade, with HSA)

Quantity 500 ml Cat. no. 12055-091 12055-083 1,000 ml
Contains L-glutamine, streptomycin sulfate at 50 g/ml, and gentamicin sulfate at 10 g/ml .
Neurobasal Medium (1X), liquid

Neurobasal Medium is for long-term growth of fetal hippocampal neurons and many other neurons of the CNS .1
AIM V Medium, liquid (research grade, with BSA)

Contains AlbuMAX supplement (bovine serum albumin) substituted for human serum albumin, L-glutamine, streptomycin sulfate at 50 g/ml, and gentamicin sulfate at 10 g/ml .
117

AIM V Media (therapeutic grade)
Performance testing: Performance tested in a growth assay using Jurkat cells (ATCC Jurkat T1B-152) . Viral testing: Raw material components of human origin are nonreactive, at the donor level, for Hepatitis B surface antigen (HBsAG), antibody to Hepatitis C virus (anti-HCV), antibodies to Human Immunodeficiency Viruses 1 and 2 (anti-HIV 1 and 2), and syphilis, as determined by currently approved testing procedures . Raw material components of human origin are also tested for alanine aminotransferase (ALT) enzyme, and found to be within normal limits . Where possible, such components are further processed by heat treatment in liquid form at 60C for 10 hours by validated methods demonstrated to inactivate adventitious viral contaminants (performed by our raw material vendors) . Intended uses: for human ex vivo tissue and cell culture processing applications .
MesenPRO RS Medium
MesenPRO RS is a reduced-serum (2%) medium specifically formulated to support the growth of human mesenchymal stem cells (MSCs) in culture . MesenPRO RS Medium consistently improves MSC expansion compared with conventional serum-supplemented media . Using MesenPRO RS Medium, MSCs can be expanded for multiple passages while maintaining their multipotential phenotype (i .e ., differentiation into osteogenic, chondrogenic, and adipogenic lineages) . MesenPRO RS consists of basal liquid medium (MesenPRO RS Basal Medium), and frozen supplement (MesenPRO RS Growth Supplement), which is added at the time of use . Applications: Formulated for the growth and expansion of mesenchymal stem cells . Recommended storage conditions: MesenPRO RS Basal Medium, 2C to 8C, in the dark; MesenPRO RS Growth Supplement, 5C to 20C, in the dark . Both the Basal Medium and Growth Supplement are guaranteed stable for 12 months when properly stored . Intended use: In vitro diagnostic (IVD) . Notes: MesenPRO RS Medium has been developed for culturing MSCs at greater than clonal densities . MesenPRO RS complete medium (basal, supplement, and glutamine) is stable for 15 days when stored at 2C to 8C, in the dark .
AIM V Medium, liquid (therapeutic grade)

Recommended storage conditions: 2C to 8C, in the dark . Intended uses: For human ex vivo tissue and cell culture processing applications . Notes: An AIM V Medium Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
Quantity 1,000 ml 10 L Cat. no. 087-0112DK 087-0112BK
Product MesenPRO RS Medium* Related products Gentamicin Reagent Solution (10 mg/ml), liquid L-Glutamine-200 mM (100X), liquid Dulbeccos Modified Eagle Medium (DMEM) low glucose (1X), liquid Dulbeccos Phosphate Buffered Saline (DPBS) without calcium, magnesium, or phenol red (1X), liquid TrypLE Express (1X), liquid, without phenol red STRO-1 (host: mouse; clone: Stro-1) (Zymed) 100 ml 100 ml 500 ml 15710-049 25030-024 11880-028 Quantity 1 kit Cat. no. 12746-012
Contains L-glutamine, 50 g/ml streptomycin sulfate, and 10 g/ml gentamicin sulfate .
500 ml 500 ml 100 l
14190-094 12604-021 39-8401
* This product is shipped in two parts, each part having separate storage requirements and separate catalogue numbers: MesenPRO RS Basal Medium (500 ml, Cat . no . 12747-010) and MesenPRO RS Growth Supplement (10 ml, Cat . no . 12748-018) . The component catalogue numbers are not available for purchase separately .
118
StemPro hESC SFM

StemPro hESC SFM, developed for human embryonic stem cell culture, is a fully-defined, serum- and feeder-free medium (SFM) specially formulated for the growth and expansion of human embryonic stem cells (hESCs) . StemPro hESC SFM has been extensively tested and proven to maintain pluripotency in a growing list of hESC lines including BG01, BG02, BG03, H1, H9, and BG01V . In addition, StemPro hESC SFM has been used to scale up production of hESCs to over 1 1010 cells . StemPro hESC SFM has been shown to support hESC growth for >50 passages without any signs of karyotypic abnormalities and to maintain the ability of hESCs to differentiate into all three germ line lineages . StemPro hESC SFM consists of StemPro hESC Supplement, DMEM/F-12 with GlutaMAX, and Bovine Serum Albumin 25% (BSA) . This product is shipped in two parts with separate storage requirements: 1 . StemPro hESC Supplement (10 ml, Cat . no . A1000601); 2 . Bovine Serum Albumin 25% (BSA) (40 ml, Cat . no . A1000801) and DMEM/F-12 with GlutaMAX (500 ml, Cat . no . 10565-018) . These components, with the exception of DMEM/F12 with GlutaMAX, are not available for purchase separately . Applications: Formulated for the growth and expansion of human embryonic stem cells . Recommended storage conditions: StemPro hESC Supplement, 5C to 20C, in the dark; DMEM/F-12 with GlutaMAX and Bovine Serum Albumin 25% (BSA), 2C to 8C (protect from light) . These components are guaranteed stable for 12 months when properly stored . Intended use: For research use only . Notes: Additional components required for complete medium: FGF BASIC (full length) REC HU (Cat . no . PHG0261), 2-Mercaptoethanol (Cat . no . 31350-010) . StemPro hESC SFM Complete Medium (Supplement, DMEM/F-12, and BSA) is stable for 1 day when stored in the dark from 2C to 8C . We recommend reconstituting minimal volumes of working complete medium (for the day), just prior to use and pre-equilibrating to temperature and gases .
Product StemPro hESC SFM Related products 2-Mercaptoethanol BG01V/hOG Cells Collagenase Type IV DPBS, without calcium, magnesium, or phenol red FGF BASIC (full length) REC HU Geltrex Reduced Growth Factor Basement Membrane Matrix SSEA1 (host: mouse; clone: MC-480) SSEA3 (host: rat; clone: MC631) SSEA4 (host: mouse; clone: MC813-70) StemPro EZPassage TRA-1-60 (host: mouse; clone: cl .A) TRA-1-81 (host: mouse; clone: cl .26) TrypLE Express Stable Trypsin Replacement Enzyme without phenol red 20 ml 1 vial 1g 500 ml 100 g 5 ml 100 g 100 g 100 g 1 box, 10 units 100 g 100 g 100 ml 31350-010 R7799-105 17104-019 14190-094 PHG0261 12760-021 41-1200 41-4400 41-4000 23181-010 41-1000 41-1100 12604-013 Quantity 1 kit Cat. no. A1000701


StemPro-34 SFM (1X), liquid
S tem Pro -34 SFM is a serum-free medium specifically formulated to support the development of human hematopoietic cells in culture .1 StemPro-34 SFM can be used to study hematopoietic cells from bone marrow, peripheral blood, or neonatal cord blood . StemPro-34 SFM is performance-tested on normal human bone marrow . It is manufactured with components of human, recombinant, or synthetic origin . StemPro-34 SFM consists of a basal liquid medium and frozen supplement (StemPro-Nutrient Supplement) which is added at the time of use . Recommended storage conditions: medium, 2C to 8C; supplement, 5C to 20C . Notes: An intended use disclosure is required for purchase of StemPro-34 SFM . For further regulatory information on the uses of this product, please contact theTech-LineSM .
KnocKout Serum Replacement

KnocKout Serum Replacement (KnocKout SR) is a defined, serum-free formulation optimized to grow and maintain undifferentiated ES cells in culture .1 It directly replaces FBS in existing protocols . The performance of KnocKout SR is enhanced when used with KnocKout DMEM . Product performance is confirmed by evaluation of its ability to support the growth of undifferentiated D3 ES cell colonies on inactivated mouse embryonic fibroblasts . Recommended storage conditions: 5C to 20C, in the dark .
Reference
1 . Goldsborough, M ., Tilkins, M . L ., Price, P ., Lobo-Alfonso, J ., Morrison, J ., Stevens, M ., Meneses, J ., Pederson, R ., Koller, B ., and Latour, A . (1998) FOCUS 20, 8 .
Related Reading References

1 . Daley, J .P ., Dadey, B .M ., Wysocki, M .G ., Caligiuri, M .A ., and Biddle, W .C . (1996) FOCUS 18, 62 . 2 . Taylor, M . et al . (2001) Blood 98, 11951199 . 3 . Szilvassy, S . et al . (1999) Blood 93, 15571566 . 4 . Delehanty, L . et al . (2003) Blood 101, 17441751 . 5 . Kirshenbaum, A . et al . (1999) Blood 94, 23332342 . 6 . Szilvassy, S ., Meyerrose, T ., and Grimes, B . (2000) Blood 95, 28292837 . 7 . Yahata, T . et al . (2003) Blood 101, 29052913 . 8 . Wang, J . et al . (2003) Blood 101, 29242931 . 9 . Kolbus, A . et al . (2003) Blood 102, 31363146 . 10 . Goyama, S . et al . (2004) Blood 104, 35583564 . 1 . Amit, M ., Carpenter, M .K ., Inokuma, M ., Chiu, C ., Harris, C .P ., Waknitz, M .A ., Itskovitz-Eldor, J . and Thomson, J .A . Clonally Derived Human Embryonic Stem Cell Lines Maintain Pluripotency and Proliferative Potential for Prolonged Periods of Culture . Developmental Biology 227, 271-278, (2000) . 2 . Kawasaki, H ., Suemori, H ., Mizuseki, K ., Watanabe, K ., Urano, F ., Ichinose, H ., Haruta, M ., Takahashi, M ., Yoshikawa, K ., Nishikawa, S ., Nakatsuji, N . and Sasai, Y . Generation of dopaminergic neurons and pigmented epithelia from primate ES cells by stromal cell-derived inducing activity . Proc. Natl. Acad. Sci. 99;3, 15801585, (2002) . 3 . Lebkowski, J .S ., Gold, J ., Xu, C ., Funk, W ., Chiu, C ., and Carpenter, M .K . Human Embryonic Stem Cells: Culture, Differentiation, and Genetic Modification for Regenerative Medicine Applications . Cancer Journal 7(2), S83-S93, (Nov-Dec 2001) . 4 . Nakayama, N ., Lee, J . and Chiu, L . Vascular endothelial growth factor synergistically enhances bone morphogenetic protein-4-dependent lymphohematopoietic cell generation from embryonic stem cells in vitro. Blood 1;95(7), 2275-2283 (2000) . 5 . Schuldiner, M ., Yanuka, O ., Itskovitz-Eldor, J ., Melton, D .A . and Benvenisty, N . Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells . Proc. Natl. Acad. Sci. USA 97;21, 11307-11312 (2000) . 6 . Amit, M ., Shariki, C ., Margulets, V . and Itskovitz-Eldor, J . (2004) Biol Reprod. 70(3), 837845 . 7 . Cheng, J ., Dutra, A ., Takesono, A ., Garrett-Beal, L . and Schwartzberg, P .L . (2004) Genesis 39, 100-104 . 8 . Rosler, E .S ., Fisk, G .J ., Ares, X ., Irving, J ., Miura, T ., Rao, M .S . and Carpenter, M .K . (2004) Dev Dyn. 229(2), 25974 . 9 . Schulz, T ., et al . (2004) Stem Cells 22, 12181238 . 10 . Wang, L ., et al . (2005) Blood Jun 15;105(12):4598-603 . Epub 2005 Feb 17 .
Quantity Cat. no.
500 ml
10639-011
DMEM/F-12 (1X), liquid, 1:1

A GlutaMAX-I medium . Recommended storage conditions: 2C to 8C . Intended uses: In vitro diagnostic (IVD) . For product formulation see page 91 .
Contains GlutaMAX-I substituted on a molar equivalent basis for L-glutamine, but no HEPES buffer .
Related products Page Geneticin Selective Antibiotic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .153 TrypLE Cell Dissociation Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .159
120
KnocKout DMEM
KnocKout DMEM is a basal medium optimized for growth of un-differentiated ES cells .1 The osmolarity is optimized to approximate that of mouse embryonic tissue . Recommended storage conditions: 2C to 8C, in the dark . Notes: Prior to use, KnocKout DMEM must be supplemented with either KnocKout Serum Replacement or FBS . Formulated without L-glutamine for increased stability; add 5 ml of 200 mM glutamine or 5 ml of GlutaMAX-I Supplement per 500 ml of medium prior to use .
Reference
1 . Goldsborough, M ., Tilkins, M . L ., Price, P ., Lobo-Alfonso, J ., Morrison, J ., Stevens, M ., Meneses, J ., Pederson, R ., Koller, B ., and Latour, A . (1998) FOCUS 20, 8 .
Quantity 500 ml
Cat. no. 10829-018
Related products Page Lipofectamine 2000 Transfection Reagent . . . . . . . . . . . . . . . . . . . . . . . . .132 TrypLE Cell Dissociation Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .159
Serum-free and Specialty Media and Reagents 121


Geneticin Selective Antibiotic
Look for consistency in these three G-418 specifications:
Purity: Purity of Geneticin selection agent is consistently greater than 90%, as determined by HPLCfar higher than any other supplier . This means, you can generally use 1530% lower concentrations of Geneticin agent than other G-418 products to achieve comparable selection results . Potency: This is one of the most misunderstood and misused criteria for comparing and assessing product quality . Potency is a prokaryotic selection parameter . Higher-potency G-418 is most effective for mammalian cell selection when it is also high in purity . Gentamicins that commonly contaminate G-418 preparations will add to potency in bacterial assays . However, they have no effect on mammalian cell selectivity . Furthermore, other bacterial contaminants in impure G-418 may be toxic to mammalian cells in culture, consequently lowering the ED50 value . ED50 Assay: This is a true measure of eukaryotic growth selectivity as determined on reference NIH3T3 cells . Higher purity generally translates to higher ED50 values . Toxic impurities lower the ED50 value, which means a more narrow working range of antibiotic selection . A consistent ED50 from a supplier assures performance reproducibility from lot to lot in mammalian cell culture applications . A selection agent commonly used for the selection of mammalian, plant, or yeast cells . This selection reagent is available in powder form or as a convenient aseptic liquid, provided at 50 mg/ ml active concentration for consistent potency . Geneticin agent is an analog of neomycin sulfate that interferes with the function of 80S ribosomes and protein synthesis in eukaryotic cells . The Tn5 or Tn601 aminoglycoside phosphotransferase gene (neo) confers resistance . When evaluating G-418 products, it is important to consider more than just potency . It is critical to look at purity and working range . Taken together, these three characteristics provide a true evaluation of the effectiveness of the G-418 preparation (sidebar) . To demonstrate the higher standards of the Geneticin agent, we recently compared three aspects of Geneticin reagent to various lots of G-418 from two different suppliers (see table below) . In all of the parameters, Geneticin reagent proved to be more consistent . Geneticin is available in both solid (powder) and liquid formats . The higher-potency liquid eliminates the time and effort spent producing working stocks from powder . Each lot is supplied at 50 mg/ml active concentration for consistent potency . Each lot is tested in an in vitro cell culture assay to ensure performance and validate label potency . Formulation: C20H40N4O102H2SO4 Formula weight: 496.6196.1 g/mol Contents and storage: Liquid Geneticin: Supplied as a 50 mg/ml solution; store at 20C, or 2C8C; guaranteed for 6 months when stored correctly . Powder Geneticin: store at room temperature prior to reconstitution, and at 2C to 8C after reconstitution; guaranteed stable for 6 months when stored correctly .
Product Geneticin Selective Antibiotic, powder Quantity 1g 5g 10 g 25 g 20 ml (50 mg/ml) 100 ml (50 mg/ml) Cat. no. 11811-023 11811-031 11811-064 11811-098 10131-019 10131-027
Geneticin Selective Antibiotic, liquid
Comparison of G-418 Specifications of Geneticin vs. Products from Suppliers A and B

Specification Purity (HPLC) Potency claimed (g/mg) Potency upon retest (g/mg) ED50 (g/ml) Invitrogen >9093% 718735 718735 2,4502,700 Supplier A 6675% 712724 640659 1,3503,100 Supplier B 6582% 673735 621677 6002,350 The Geneticin advantage Significantly higher purity Most stringent potency assay Consistently higher actual potency Reliable selection, lot to lot
Comparison of Geneticin specifications with alternative G-418 suppliers . Multiple lots of G-418 from two different suppliers were analyzed according to Invitrogen raw material QC protocols for Geneticin . Results obtained with G-418 from Supplier A and Supplier B displayed wide variation on all measured parameters . Discrepancies between retested potency values and those claimed by other suppliers may be due to differences in G-418 reference lots and assay procedures .
122
Blasticidin S HCl, powder

Blasticidin is a nucleoside antibiotic from Streptomyces griseochromogenes . It is a potent translational inhibitor in both prokaryotic and eukaryotic cells . Resistance to blasticidin is conferred by the bsd gene product from Aspergillus terreus . Blasticidin has a fast, potent mode of action, causing rapid cell death at low antibiotic concentrations . E. coli strains are generally sensitive to concentrations of 50 g/ml, while mammalian cells are sensitive to concentrations as low as 2 to 10 g/ml . Cell death occurs rapidly, and blasticidin-resistant stable mammalian cell lines can be generated in less than one week, at low concentrations . Recommended storage conditions: 20C .
Fetal Bovine Serum, ES cell-qualified

Origin: United States
Specially tested for the ability to sustain undifferentiated cellular morphology of embryonic stem cells . Endotoxin level: 50 EU/ml . Hemoglobin level: 25 mg/dl . Recommended storage conditions: 5C to 20C . Intended use: In vitro diagnostic (IVD) .
Quantity 100 ml Cat. no. 16141-061 16141-079 500 ml
Quantity 50 mg Cat. no. R210-01
Hygromycin B
Hygromycin B is an aminoglycosidic antibiotic that inhibits protein synthesis by disrupting translocation and promoting mistranslation at the 80S ribosome . Because it uses a different mode of action than Geneticin, blasticidin S, or Zeocin, its perfect for dual-selection experiments when used in conjunction with another selection agent . Provided in solution at a concentration of 50 mg/ml . Recommended storage conditions: 4C .
Origin: Countries meeting USDA importation requirements

Specially tested for the ability to sustain undifferentiated cellular morphology of embryonic stem cells . Endotoxin level: 50 EU/ml . Hemoglobin level: 25 mg/dl . Recommended storage conditions: 5C to 20C . Intended use: In vitro diagnostic (IVD) .
Quantity 100 ml Cat. no. 10439-016 10439-024 500 ml
GlutaMAX-I Supplement
Extremely stable in aqueous solution . See Appendix I, page 202 . 200 mM supplied in 0 .85% NaCl . Recommended storage conditions: 5C to 20C .
CO2-Independent Medium (1X), liquid
Reference
1 . Christie, A . and Butler, M . (1994) FOCUS 16, 9 .
A non-HEPES proprietary medium suitable for supporting cell growth for a variety of epithelial, fibroblast, and lymphoid cell lines in atmospheric conditions .1 It is ideal for transporting cells or tissue and for the handling of mouse embryos in atmospheric conditions . Not intended for CO2-based mouse embryo culture . Recommended storage conditions: 2C to 8C, in the dark .
Contains the dipeptide L-alanyl-L-glutamine .
Reference
1 . Battista, P .J . and Weiss, S .A . (1991) FOCUS 13, 110 . Cat. no. 35050-038 35050-087
Quantity 500 ml 10 500 ml
Cat. no. 18045-054 18045-070
TrypLE Express, Stable Trypsin Replacement Enzyme (1X), liquid

A pure enzyme for gentler cell dissociationideal for stem cells . This recombinant enzyme is an alternative to animal trypsin for the dissociation of adherent mammalian cells . See page 151 for ordering information .
123

Basic Fibroblast Growth Factor (bFGF), Human, Recombinant
bFGF, Human, Recombinant, also known as Heparin Binding Growth Factor (HBGF)-2, is a 17kDa protein supplied lyophilized with no additives . Basic FGF is structurally similar to acidic FGF, but is functionally different . Applications: Studies of angiogenesis; mitogenesis of fibroblasts; neurite outgrowth in PC-12 cells; receptor binding; tyrosine protein kinase phosphorylation . Performance and quality testing: Determined to be >95% pure by SDS-PAGE and N-terminal sequencing . Functional performance is verified by thymidine incorporation into serum-starved Balb/c 3T3 cells . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in dilute solution . Membrane-filtered through a 0 .22 m filter .
Platelet-derived Growth Factor BB (PDGF BB), Human, Recombinant

PDGF BB, Human, Recombinant, is supplied lyophilized from water . Applications: Receptor binding studies; studies of transmembrane signaling and protein phosphorylation; and fibroblast proliferation studies . Performance and quality testing: PDGF BB is determined to be >97% pure by SDS-PAGE . Functional performance is verified by thymidine incorporation into confluent Balb/c 3T3 cells . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in glass or in dilute solution . Membrane-filtered through a 0 .22 m filter .
Quantity 100 g 1mg 10 g 25 g PDGF-BB Recombinant Mouse PDGF-BB Recombinant Mouse PDGF-BB Recombinant Mouse PDGF-BB Recombinant Mouse Cat. no. PMG0041 PMG0043 PMG0044 PMG0045
Quantity 100 g 10 g 25 g 100 g EGF Recombinant Human EGF Recombinant Human EGF Recombinant Human EGF Recombinant Human (Liquid Form) Cat. no. PHG0311 PHG0314 PHG0315 PHG0311L
Vascular-Endothelial Products
Human Endothelial-SFM
Human Endothelial-SFM is a basal serum-free growth medium developed to support the isolation and expansion of human umbilical venous and arterial endothelial cells .14 Medium requires supplementation with bFGF (20 ng/ml), EGF (10 ng/ml), and human plasma fibronectin (10 g/ml) . Recommended storage conditions: 2C to 8C, in the dark . Notes: A Human Endothelial-SFM Master File has been submitted to the FDA . Permission to cross-reference the Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
Insulin-like Growth Factor-I (IGF-I), Human, Recombinant

IGF-I, Human, Recombinant, is prepared from E. coli and supplied lyophilized from water . Applications: Receptor binding, transmembrane signaling, protein phosphorylation, and epidermal cell proliferation studies . Performance and quality testing: Determined to be >97% pure by SDS-PAGE . Functional performance is verified by thymidine incorporation into Balb/c 3T3 cells . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in glass or in dilute solution . Membrane-filtered through a 0 .22 m filter .
Quantity 10 g Cat. no. PHG9074
124
References
1 . 2 . 3 . 4 . Battista, P .J ., Bowen, H .J ., and Gorfien, S .F . (1995) FOCUS 17, 10 . Soderland, C ., Veres, J .S ., and Battista, P .J . (1995) FOCUS 17, 14 . Battista, P .J . and Soderland, C . (1995) FOCUS 17, 106 . Battista, P .J ., Soderland, C ., Tilkins, M .L ., and Gorgien, S .F . (1996) American Biotechnology Laboratory 14, 34 . 5 . Battista, P .J . and Soderland, C . (1998) Cell Biology: A Laboratory Handbook 1, 137 .
Fibronectin, Human Plasma

Fibronectin is a large cell-surface and plasma protein that exhibits structural and adhesive properties in cell-associated fibrillar matrices . Fibronectin is one of the primary cell adhesion molecules . Human fibronectin is filtered (0 .2 m) and supplied lyophilized . Performance and quality testing: Purity is determined by SDS-PAGE . Human fibronectin is assayed for its ability to support cell attachment and spreading, and screened to confirm absence of mycoplasmas or bacterial contamination . Recommended storage conditions: 5C to 20C . Notes: Do not store in dilute solution . Avoid repeated freezing and thawing .
Quantity 500 ml
Cat. no. 11111-044
Basic Fibroblast Growth Factor (bFGF), Human, Recombinant

bFGF, Human, Recombinant, also known as Heparin Binding Growth Factor (HBGF)-2, is a 17kDa protein supplied lyophilized with no additives . Basic FGF is structurally similar to acidic FGF, but is functionally different . Applications: Studies of angiogenesis; mitogenesis of fibroblasts; neurite outgrowth in PC-12 cells; receptor binding; tyrosine protein kinase phosphorylation . Performance and quality testing: Determined to be >95% pure by SDS-PAGE and N-terminal sequencing . Functional performance is verified by thymidine incorporation into serum-starved Balb/c 3T3 cells . Recommended storage conditions: 20C . Notes: Avoid repeated freezing and thawing . Do not store in dilute solution . Membrane-filtered through a 0 .22 m filter .
Quantity 5 mg Cat. no. 33016-015
125
Cytogenetics Products
Quality Control
All MarrowMAX, AmnioMAX, and KaryoMAX products have been stringently qualified by an independent, clinically certified cytogenetic laboratory using standardized cytogenetic protocols . This qualified line of products helps you attain reproducible, quality results in standard clinical cytogenetic protocols involving the analysis of either amniotic fluid cells, chorionic villus samples, peripheral blood lymphocytes, or bone marrow cells . Warning: Although the media and reagents are safe to use, the human peripheral blood used in procedures with these products may be ` infectious or hazardous to the user . Be sure to use proper handling, decontamination, and waste disposal techniques .
MarrowMAX Bone Marrow Medium

MarrowMAX Bone Marrow Medium is a fully supplemented medium developed specifically to support bone marrow and peripheral blood cell culture for in vitro cytogenetic analysis of hematological disease . Recommended storage conditions: 5C to 20C, in the dark . Intended use: In vitro diagnostic (IVD) .
This product is the subject of claims in US Patent No . 6,103,522 and equivalent foreign patents owned by the Fred Hutchinson Cancer Research Center . Purchase of this product conveys to the buyer only the non-transferable right under these patents to use the purchased amount of the product in research conducted by the buyer . The buyer cannot sell or otherwise transfer this product or any derivatives thereof to any third party . The buyer may not use this product or any derivatives thereof for any in vivo diagnostic or clinical use in humans . Fred Hutchinson Cancer Research Center and Invitrogen Corporation reserve all other rights, and this product may not be used in any manner other than as provided herein . In particular, without prior written agreement between the buyer and Invitrogen, no rights under the above patents are conveyed to the buyer to use the product in manufacturing, or to resell the product or components of the product as a stand-alone reagent or in a kit, whether or not such stand-alone reagent or such kits are resold for use in research . If the purchaser is not willing to accept the limitations of this limited use label license, Invitrogen is willing to accept return of the product with full refund . For information on purchasing a license to this product for purposes other than research, contact the Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N . C2M-027, Seattle, WA 98104 if such use is in vivo diagnostic or clinical use in humans, or Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, CA 92008 . Phone 760 .603 .7200 . Fax 760 .602 .6500 if such use is for the manufacture of a product for use in an ex vivo diagnostic or as a pharmaceutical product .
MarrowMAX, AmnioMAX, and KaryoMAX ProductsCertified for Consistency in Cytogenetics

MarrowMAX, AmnioMAX, and KaryoMAX products are specifically formulated by us and application tested by an independent laboratory to deliver clear, reproducible results in standard clinical cytogenetic protocols involving the analysis of amniotic fluid cells, chorionic villus samples, peripheral blood lymphocytes, or bone marrow cells . The combination of the gibco production process and strict quality control provides you with confidence that every lot of MarrowMAX, AmnioMAX, or KaryoMAX products will support healthy cell morphology and rapid culture growth . Our goal is to help you obtain results that are easy to analyze and interpret .
Quality Control Testing

Product MarrowMAX Bone Marrow Medium AmnioMAX-II Complete Medium AmnioMAX-C100 Complete Medium (system) AmnioMAX-C100 Basal Medium AmnioMAX-C100 Supplement PB-MAX Karotyping Medium KaryoMAX Colcemid Solution in PBS KaryoMAX Colcemid Solution in HBSS KaryoMAX Giemsa Stain Stock Solution Phytohemagglutinin
Colcemid is a trademark of CIBA Limited .
Base cat. no. 12260 11269 12558 17001 12556 12557 15212 15210 10092 10576
Bone marrow
Peripheral blood
Primary amniocytes
Number of colonies
Mitotic colony
Mitotic index
Banding resolution
126
AmnioMAX Complete Media

AmnioMAX media are specifically formulated for the in vitro culture of human primary amniotic fluid cells . They are designed to optimize cell attachment and growth rates, and to provide high metaphase yields . Two different formulations are available, offering cytogenetics laboratories the ability to use a secondary cell culture medium to meet established laboratory protocols . AmnioMAX-C100 Complete Medium consists of two components (basal medium and supplement), while AmnioMAX-II Complete Medium is a single -bottle formulation for added convenience .
KaryoMAX Products
We offer a full line of karyotyping products designed to minimize setup time and to maximize lab efficiency . KaryoMAX medium is fully supplemented and specifically formulated to optimize cell growth, reducing culture time and days to harvest, and the mitotic index and banding resolution .
PB-MAX Karyotyping Medium

PB-MAX Karyotyping Medium is a fully supplemented, ready-to-use RPMI-based medium designed for cultivation of lymphocytes from peripheral blood . It is performance tested on cells from whole blood . Every lot of PB-MAX Medium comes with a Certificate of Analysis detailing the mean mitotic index and mean banding resolution of chromosome 10 . Recommended storage conditions: 5C to 20C . Intended use: In vitro diagnostic (IVD) .
AmnioMAX-II Complete Medium

AmnioMAX-II Complete Medium is a second-generation formulation further optimized for cell attachment and increased pH stability .1 Supplied in a single bottle, it is complete with antibiotics and growth-promoting agents, and requires no further supplementation . Recommended storage conditions: 5C to 20C, in the dark . Intended use: In vitro diagnostic (IVD) .
Quantity 100 ml 500 ml
Contains fetal bovine serum, L-glutamine, and phytohemagglutinin .
Cat. no. 12557-013 12557-021
Reference
1 . Kuligowski, S ., Csirke, B ., Biddle, W .C ., Battista, P ., and Kielma, M . (1999) FOCUS 21, 35 .
KaryoMAX Colcemid Solution, liquid (10 g/ml), in HBSS

KaryoMAX Colcemid Solution is functionally tested as a blocking agent that arrests cells in metaphase by disrupting the cytoskeleton . It is performance tested on cells from whole blood . Every lot comes with a Certificate of Analysis detailing the mean mitotic index . Prepared in Hanks Balanced Salt Solution . Recommended storage conditions: 2C to 8C . Intended use: In vitro diagnostic (IVD) .
AmnioMAX-C100 Products
The system consists of AmnioMAX-C100 Basal Medium and AmnioMAX-C100 Supplement . It is complete with antibiotics and growth-promoting agents, and requires no further supplementation .1 The medium and supplement are also available separately . Recommended storage conditions: medium, 2C to 8C, in the dark; supplement, 5C to 20C . Intended use: In vitro diagnostic (IVD) .
Reference
1 . Biddle, W .C ., Kuligowski, S ., Lockwood, D .H ., Filby, J ., and Custer-Hagen, T . (1992) FOCUS 14, 80 .
Product AmnioMAX-C100 Complete Medium, system AmnioMAX-C100 Basal Medium, liquid AmnioMAX-C100 Supplement, liquid Quantity 1 set 90 ml 450 ml 15 ml 75 ml Cat. no. 12558-011 17001-082 17001-074 12556-015 12556-023
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KaryoMAX Colcemid Solution, liquid (10 g/ml), in PBS

KaryoMAX Colcemid Solution is functionally tested as a blocking agent that arrests cells in metaphase by disrupting the cytoskeleton . It is performance tested on cells from whole blood . Every lot comes with a Certificate of Analysis detailing the mean mitotic index . Prepared in phosphate-buffered saline, but with no phenol red . Recommended storage conditions: 2C to 8C . Intended use: In vitro diagnostic (IVD) .
Related Products for Cytogenetics

Gurr Buffer Tablets (pH 6.8)
Gurr Buffer Tablets offer a convenient way to prepare a liter of buffer at pH 6 .8 for dilution of Giemsa stain . Simply add a liter of distilled water to a Gurr Buffer Tablet and stir until the tablet is dissolved .
Phytohemagglutinin (M Form) (PHA), liquid

Phytohemagglutinin is a liquid extract of the red kidney bean Phaseolus vulgaris . This extract is capable of inducing blastogenesis in vitro in various mammalian mononuclear cells . Concentration: Because of the undefined nature of this extract, we are not able to assign weight/volume values on the active material in our final product . Performance and quality testing: Each lot is assayed in a performance test system . Recommended storage conditions after rehydration: 5C to 20C . Attention: This material is hazardous . All persons using this product should review the Material Safety Data Sheet before handling .
KaryoMAX Giemsa Stain Stock Solution

Functionally tested for consistent G-banding of chromosomes . Intended use: In vitro diagnostic (IVD) .
Potassium Chloride Solution, 0.075 M

A premixed 0 .075 M hypotonic solution . Enlarges cells and facilitates the spreading of metaphase chromosomes for subsequent analysis . Recommended storage conditions: 2C to 8C . Intended use: In vitro diagnostic (IVD) .
Quantity 4 100 ml (case) Cat. no. 10575-090
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Antibiotic-Antimycotic (100X), liquid

Spectrum: Bacteria, fungi, and yeasts . Recommended storage conditions: 5C to 20C . Attention: This material is hazardous . All persons using this product should review the Material Safety Data Sheet before handling .
Penicillin-Streptomycin, liquid
Spectrum: Gram-positive and gram-negative bacteria . Recommended storage conditions: 5C to 20C . Attention: This material is hazardous . All persons using this product should review the Material Safety Data Sheet before handling .
Contains 10,000 units of penicillin (base), 10,000 g of streptomycin (base), and 25 g of amphotericin B (Fungizone Antimycotic)/ml in 0 .85% saline .
Contains 10,000 units of penicillin (base) and 10,000 g of streptomycin (base) per ml, utilizing penicillin G (sodium salt) and streptomycin sulfate in 0 .85% saline .
Fungizone Antimycotic, liquid

Spectrum: Fungi and yeasts . Recommended concentration: 0 .25 to 2 .5 g/ml . Recommended storage conditions: 5C to 20C . Attention: This material is hazardous . All persons using this product should review the Material Safety Data Sheet before handling .
Quantity 100 ml
Cat. no. 15070-063
Contains 5,000 units of penicillin (base) and 5,000 g of streptomycin (base) per ml, utilizing penicillin G (sodium salt) and streptomycin sulfate in 0 .85% saline .
Contains 250 g of amphotericin B and 205 g of sodium deoxycholate per ml as a solubilizer in distilled water .
Gentamicin Reagent Solution (10 mg/ml), liquid

Spectrum: Gram-positive and gram-negative bacteria . Recommended concentration: 0 .5 to 50 g/ml . Recommended storage conditions: 15C to 30C . Attention: This material is hazardous . All persons using this product should review the Material Safety Data Sheet before handling .
Contains 10 mg/ml gentamicin sulfate in distilled water .
Cat. no. 15710-064 15710-072
Contains 50mg/ml gentamicin sulfate in distilled water .
Fungizone is a trademark of E .R . Squibb & Sons, LLC .
Cat. no. 15750-060 15750-078
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Collagenase, lyophilized (Clostridiopeptidase A)

Prepared from Clostridium histolyticum . E.C. Number: 3 .4 .2 .4 .3 . Activity: 125 to 200 units/mg weight . One unit will liberate 1 mol amino acid from collagen, expressed as L-leucine, in 18 hr at 37C . Recommended storage conditions: 2C to 8C .
RPMI Medium 1640 (1X), liquid (Folate-free)

Quantity 500 ml
Contains L-glutamine, but no folic acid .
Cat. no. 27016-021
Sodium Bicarbonate Solution, 7.5% (w/v)

200 mM, supplied in 0 .85% NaCl . Extremely stable in aqueous solution . See Appendix I, page 202 . Recommended storage conditions: 5C to 20C .
Quantity 100 ml 20 x 100 ml Cat. no. 25080-060 25080-102
Reference
1 . Christie, A . and Butler, M . (1994) FOCUS 16, 9 .
Water, distilled
Membrane filtered and endotoxin screened . Recommended storage conditions: 15C to 30C .
Cat. no. 35050-061 35050-079
Contains the dipeptide L-alanyl-L-glutamine .
Quantity 100 ml 20 100 ml (case) 500 ml 10 500 ml (case) 1,000 ml Cat. no. 15230-071 15230-196 15230-162 15230-788 15230-147
L-Glutamine-200 mM (100X), liquid

Supplied at 29 .2 mg/ml in 0 .85% NaCl . pH: 4 .7 to 6 .0 . Recommended storage conditions: 5C to 20C . Intended use: In vitro diagnostic (IVD) .
130
Overview of Transfection, Selection, and Protein Delivery Methods

The easy, fast, and reliable introduction of DNA, siRNA, RNA, and protein into mammalian cells is crucial to experimental results . Improvements in transfection reagent technology have allowed researchers to quickly discover novel genes and rapidly elucidate gene function . Transfection: Reagent-mediated transfection is a fast, simple, and reproducible means for easily introducing DNA, RNA, siRNA, or oligonucleotides into eukaryotic cells (Figure 1) . Transfections using reagents such as Lipofectamine LTX, FreeStyle MAX, and Lipofectamine 2000 are superior to older methods of transfection like DEAE-dextran and calcium phosphate precipitation for both transient protein expression and the generation of high-expression stable cell lines . Life Technologies transfection reagents offer consistent and reliable high performance . These reagents allow highly efficient transfection of a broad range of cell types, including adherent, suspension, insect, and primary cultures (Table 1) . All of Invitrogens transfection reagents include streamlined protocols for maximum convenience and ease of use . Many reagents include protocols for high-throughput use . Selection agents: Transfection is used in conjunction with an appropriate selection agent, i .e ., Geneticin, hygromycin, blasticidin, or Zeocin . Protein translocation: With Voyager technology, active fusion proteins can be translocated across the eukaryotic cell membrane into cells . Voyager technology is based on the properties of the herpes simplex viral protein VP22 . Using Voyager translocation, you can study protein function in cells that are extremely difficult to transfect . Voyager translocation results in a 100% homogeneous population because all of the cells will contain the translocated protein . The translocated protein can be studied almost immediately after it enters the cell . Fusion proteins can be generated in eukaryotic cells or E. coli .
Table 1Guidelines for selecting a transfection reagent.
Transfection application Most adherent cells Animal originfree RNAi Oligonucleotides (antisense) Cells plated at low density Suspension cells Suspension 293 cells Suspension CHO cells Insect cells Endothelial cells Difficult-to-transfect cells Primary cells Recommended reagent Lipofectamine 2000 Reagent Lipofectamine LTX with PLUS Reagent Lipofectamine 2000 CD Reagent FreeStyle MAX Reagent Lipofectamine 2000 Reagent Lipofectamine RNAiMAX Reagent Oligofectamine Reagent Lipofectamine LTX with PLUS Reagent FreeStyle MAX Reagent Lipofectamine LTX with PLUS Reagent FreeStyle MAX Reagent 293fectin Reagent FreeStyle MAX Reagent Cellfectin Reagent Lipofectin LTX with PLUS Reagent Lipofectamine LTX with PLUS Reagent Lipofectamine LTX with PLUS Reagent
+
+ + +
+
+
+
+ +
+
Cationic lipid reagent
+ + + +
+
+
+
+ +
+
++
+ ++ + + +
+
+ + +
+
+
+
+ +
+
+
+
+
+ +
+
+ + +
DNAcationic lipid complex
Figure 1Introduction of DNA into mammalian cells. For cell linespecific transfection protocols, visit the Cell Lines database at www .invitrogen .com/transfection .
DNA
+ +
+ +
+ +
+ +
+ +
+ ++ ++ ++
+ ++ + ++
+ + +
+ +
+ + + +
++
+ +
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+ ++ + +
N
+ + + +
++
+ + + +
++
Mammalian cell
Protein is produced
131
Transfection Reagents
Lipofectamine 2000 Transfection Reagent is a proprietary cationic lipid formulation that offers the highest transfection efficiencies on the widest variety of cell lines with high protein expression levels or gene knockdown activity (RNAi) . It is the leading transfection reagent for simple, effective, and optimal nucleic acid delivery in a wide range of eukaryotic cell types and is proven to work in high-throughput applications . Recommended storage conditions: 2C to 8C .
Lipofectamine LTX Transfection Reagent
Quantity 0 .75 ml 1 .5 ml 15 ml Cat. no. 11668-027 11668-019 11668-500
Lipofectamine LTX Reagent offers a new, advanced solution for gene expression studies in hard-to-transfect cells . No other plasmid DNA specific transfection reagent can match the efficiency, convenience, and gentleness of Lipofectamine LTX Reagent for transfection of primary, difficult-to-transfect, and sensitive cell lines . Lipofectamine LTX Reagent is synthesized from 100% animal originfree components, making it easy to validate the absence of zoonotic diseases, such as BSE or viruses, in experiments or cell lines . Lipofectamine LTX Reagent offers high transfection efficiency with maximal (>90%) cell viability using rapid, simple protocols that obviate the need for changing media after transfection . PLUS reagent enhances transfection efficiency of Lipofectamine LTX reagent in primary and hard-to-transfect cells . Recommended storage conditions: 2C to 8C .
Lipofectamine 2000 CD Transfection Reagent
Lipofectamine 2000 CD (Chemically Defined) Transfection Reagent is designed for applications where concerns about animal-derived components exist . It is synthesized from 100% animal originfree components, making it easy to validate the absence of zoonotic diseases, such as BSE or viruses, in experiments or cell lines . Lipofectamine 2000 CD Reagent is appropriate for all transfections from preclinical or academic research to large-scale bioprocess applications . Recommended storage conditions: 2C to 8C . Notes: A Drug Master File (DMF) describing the manufacture of Lipofectamine 2000 CD has been filed with to the FDA . The DMF details the process to manufacture Lipofectamine 2000 CD using animal originfree components, simplifying the FDA documentation required for your downstream product . Permission to cross-reference the Drug Master File may be obtained by contacting Invitrogen Technical Support or your local Sales Representative .
FreeStyle MAX Reagent
Generating milligram quantities of functional protein in mammalian cells has been a big challenge to pharmaceutical and preclinical researchers . The new FreeStyle MAX Reagent is a proprietary cationic lipid formulation that allows rapid generation of milligram quantities of recombinant protein in suspension CHO and 293 cells . Combining transient transfection with suspension culture protocols reduces the time required to generate milligram levels of protein from several months to one week . Protein production in CHO cells produces proteins with glycosylation patterns required for many downstream preclinical and therapeutic applications . Recommended storage conditions: 2C to 8C .
Quantity 1 ml 15 ml 10 x 15 ml Cat. no. 16447-100 16447-500 16447-750
Related products Page OptiPRO SFM (1X) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .180
Related products Page FreeStyle 293 Expression Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .177 FreeStyle 293 Expression System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .178 FreeStyle 293-F Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .177
132
293fectin Transfection Reagent

293fectin Transfection Reagent is a specially designed cationic lipid formulation that offers extremely high transfection efficiencies of FreeStyle 293-F suspension cells in serum-free Gibco FreeStyle 293 Expression Medium . It is available separately or as part of the FreeStyle 293 Expression System . Recommended storage conditions: 2C to 8C .
PLUS Reagent
PLUS Reagent was specifically designed to expand the array of transfectable cell types . Some customers find when using PLUS Reagent, less optimization of DNA concentration is needed to achieve high transfection performance . PLUS Reagent used with Lipofectamine LTX Reagent has been demonstrated to improve the transfection of BHK-21, NIH 3T3, and other cell lines .1 Recommended storage conditions: 2C to 8C .
Quantity 1 ml 15 ml 10 x 15 ml Cat. no. 12347-019 12347-500 12347-750
Reference
1 . Fawcett, T .W . and Hawley-Nelson, P . (1997) FOCUS 19, 5 .
Quantity 0 .85 ml Cat. no. 11514-015
Lipofectamine RNAiMAX Transfection Reagent

Lipofectamine RNAiMAX Transfection Reagent offers an advanced, efficient solution for siRNA delivery . No other siRNA-specific transfection reagent provides such easy and efficient siRNA delivery in a wide variety of cell lines . High knockdown levels of target genes can be achieved with as little as 1 nM siRNA . Lipofectamine RNAiMAX Transfection Reagent gives maximal knockdown and excellent cell viability across a 10-fold concentration range of the reagent . This makes Lipofectamine RNAiMAX reagent easy to optimize so results are easy to obtain in every experiment .
Lipofectamine Transfection Reagent
Lipofectamine Reagent is formulated for the transfection of DNA into eukaryotic cells and has been shown to work well with cells such as BHK-21 and NIH 3T3 . Recommended storage conditions: 2C to 8C .
Quantity 0 .75 ml 1 .5 ml Cat. no. 13778-075 13778-150
Quantity 0 .5 ml 1 ml 4 ml Cat. no. 18324-111 18324-012 18324-020
Cellfectin II Transfection Reagent

Recommended storage conditions: +4C to 8C .

Lipofectamine 2000 Reagent delivers DNA with excellent transfection performance for protein expression, gene silencing and functional assays . Superior efficiency, optimized protocols, and unparalleled success across a broad range of cell lines (figure 1) - Lipofectamine 2000 Reagent does it all . Recommended storage conditions: 2C to 8C .
Quantity 0 .75 ml 1 .5 ml 15 ml Cat. no. 11668-027 11668-019 11668-500
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Notes:
134
Notes:

Notes:
136
Chapter 5 Balanced Salt Solutions

Balanced Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138 Dulbeccos Phosphate-Buffered Salines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138 Dulbeccos Phosphate-Buffered Saline (DPBS) (1X), liquid. . . . . . . . . . . . . . . . . . . . . . . . . . 138 Dulbeccos Phosphate-Buffered Saline (DPBS) (10X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . 139 Dulbeccos Phosphate-Buffered Saline (DPBS), powder . . . . . . . . . . . . . . . . . . . . . . . . . . . 139 Earles Balanced Salt Solution. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139 Earles Balanced Salt Solution (EBSS) (1X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139 Hanks Balanced Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Hanks Balanced Salt Solution (HBSS) (1X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Hanks Balanced Salt Solution (HBSS) (10X), liquid. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140 Phosphate-Buffered Saline (PBS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Phosphate-Buffered Saline (PBS) 7.2 (1X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Phosphate-Buffered Saline (PBS) 7.2 (10X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Phosphate-Buffered Saline (PBS) 7.4 (1X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Phosphate-Buffered Saline (PBS) 7.4 (10X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Balanced Salt Solutions Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142 Balanced Salt Solutions Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
137
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Balanced Salt Solutions

For information on classifications, production, or preparation of the products in this chapter, see page 142.
Liquid balanced salt solutions

Quality Control
To ensure that our validated systems and controls continue to be effective, we confirm the quality of our balanced salt solutions by testing samples from each production batch. Chemical checks. We measure osmolality and pH of all batches to verify compliance with product specifications. Microbiological tests. For balanced salt solutions, we confirm the absence of bacterial and fungal contamination using the method referenced in the current edition of the U.S. Pharmacopeia (USP) sterility test monograph. Stability tests. The expiration date shown on every in vitro diagnostic (IVD)-labeled product is an indication of how long the product will deliver effective performance. Our stability program establishes and verifies product expiration dates. Product groups are monitored at periodic intervals to determine the length of time that the product will deliver acceptable performance under recommended storage conditions. Endotoxin tests. We determine endotoxin levels in all balanced salt solutions using a Limulus Amoebocyte Lysate (LAL) test. Salt solutions with the lowest level of endotoxin attainable in the industry today are routinely available from Invitrogen.
Powdered balanced salt solutions

Quality Control
We confirm the quality of our powdered balanced salt solutions by evaluating samples from each batch as required by specifications. Samples are prepared in 1X liquid form and membrane filtered, and testing may include: 1. Endotoxin 2. Solubility 3. Osmolality 4. pH The values for these parameters are determined using the test conditions described for liquid balanced salt solutions. All powdered balanced salt solutions are produced without sodium bicarbonate to increase stability.
Dulbeccos Phosphate-Buffered Salines

All gibco Dulbeccos Phosphate-Buffered Salines contain potassium chloride, monobasic potassium phosphate, sodium chloride, and dibasic sodium phosphate; some formulations may contain additional components. For product formulation please refer to the tables at the end of this chapter.
Dulbeccos Phosphate-Buffered Saline (DPBS) (1X), liquid

Quantity 500 ml 10 500 ml (case) 1,000 ml 6 1,000 ml (case) 5L 10L Cat. no. 14040-083 14040-091 14040-174 14040-117 14040-224 20L
Contains no calcium, magnesium, or phenol red.
Cat. no. 14190-094 14190-169 14190-136 14190-326 14190-342 14190-359 14190-367
Quantity 100 ml 500 ml 10 500 ml (case) 1,000 ml 6 1,000 ml (case)
Contains calcium and magnesium, but no phenol red.

Quantity 500 ml 1,000 ml Cat. no. 14287-080 14287-072
Contains 1,000 mg/L D-glucose and 36 mg/L sodium pyruvate, calcium, and magnesium, but no phenol red.
138
Dulbeccos Phosphate-Buffered Saline (DPBS) (10X), liquid

pH: The pH of the 10X solution is 4.4 to 4.7. The pH of the 1X solution will be acidic after 1:10 dilution and must be adjusted to 7.1 0.1. Recommended storage conditions: 15C to 30C. Intended use: In vitro diagnostic (IVD).
Dulbeccos Phosphate-Buffered Saline (DPBS), powder

Quantity 1 10 L Cat. no. 21600-069 21600-010 21600-044 10 1 L 1 50 L
Contains no calcium or magnesium.
Quantity 500 ml
Cat. no. 14080-048
pH: The pH of the 10X solution is 6.7 to 7.0. The pH of the 1X solution should be checked after 1:10 dilution to ensure that it is 7.1 0.1. If the pH is not 7.1 0.1, adjust as outlined in Balanced Salt Solutions Preparation, page 123.
Contains no calcium or magnesium.
Quantity 1 10 L 11L
Cat. no. 21300-058 21300-017
Contains magnesium. Calcium chloride is supplied separately in an amount sufficient for 0.1 g/L.
Cat. no. 14200-067 14200-083
Comparison of Dulbeccos Phosphate-Buffered Salines (DPBS)

Base cat. no. With 1,000 mg/L D-glucose and 36 mg/L sodium pyruvate Without calcium chloride and magnesium chloride Without phenol red 10X concentration Powder
* Standard formulation, as published in (1954) J. Exp. Med. 99, 167. Powdered formulations of DPBS, CaCl2 is supplied separately at 0.1 g/L. Use this comparison table as a quick reference for identifying the base catalogue number of the product thats best for your application. For the exact composition of each product, see the formulation tables on page 115.
14040*
14287
14080
21300
14190
14200
21600
Earles Balanced Salt Solution

Earles Balanced Salt Solution (EBSS) (1X), liquid

Quantity 500 ml
Contains no calcium, magnesium, or phenol red.
Cat. no. 14155-048
139
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Hanks Balanced Salt Solutions

All gibco Hanks Balanced Salt Solutions contain potassium chloride, monobasic potassium phosphate, sodium chloride, dibasic sodium phosphate, and D-glucose; some formulations may contain additional components. For product formulation please refer to the tables at the end of this chapter.
Hanks Balanced Salt Solution (HBSS) (1X), liquid

Hanks Balanced Salt Solution (HBSS) (10X), liquid

Quantity 500 ml
Contains calcium and magnesium.
Cat. no. 24020-091 Quantity 500 ml Cat. no. 14185-045
Contains no calcium chloride, magnesium chloride, magnesium sulfate, sodium bicarbonate, or phenol red.
Quantity 500 ml 1,000 ml 10 500 ml (case) 6 1,000 ml (case)

Cat. no. 14025-050 14025-076 14025-100 14025-126
Quantity 500 ml
Cat. no. 14065-049
Contains calcium and magnesium, but no sodium bicarbonate or phenol red.
Quantity 100 ml 500 ml 10 500 ml (case) 5L 10L
Cat. no. 14170-070 14170-088 14170-138 14170-195 14170-203
Contains no calcium chloride, magnesium chloride, or magnesium sulfate.
Quantity 500 ml 10 500 ml (case) 1,000 ml 6 1,000 ml (case)
Cat. no. 14175-053 14175-129 14175-079 14175-145
Contains no calcium chloride, magnesium chloride, magnesium sulfate, or phenol red.
Comparison of Hanks Balanced Salt Solutions (HBSS)

Base cat. no. Without calcium chloride, magnesium chloride, or magnesium sulfate Without phenol red 10X concentration 24020* 14025 14065 14170 14175* 14185
* Standard formulation, as published in Hanks, J.H. and Wallace, R.E. (1949) Proc. Soc. Exp. Biol. Med. 71, 196, ModificationNational Institutes of Health. MgCl26H2O added to original formulation. Original formulation calls for 0.06 g/L Na2PO 42H2O and 0.02 g/L phenol red. Hanks Balanced Salt Solutions are intended for use in ambient air, not in CO2-equilibrated incubators. Use this comparison table as a quick reference for identifying the base catalogue number of the product thats best for your application. For the exact composition of each product, see the formulation tables on page 127.
140
Phosphate-Buffered Saline (PBS)

All GIBCO Phosphate-Buffered Salines contain monobasic potassium phosphate, sodium chloride, and dibasic sodium phosphate; some formulations may contain additional components. For the exact composition of each product, see the formulation tables on page 144. For product formulation please refer to the tables at the end of this chapter.
Phosphate-Buffered Saline (PBS) 7.2 (1X), liquid

pH: 7.2 0.1 Intended use: In vitro diagnostic (IVD).

pH: 7.4 0.1 Intended use: In vitro diagnostic (IVD).
Quantity 500 ml 10 500 ml (case) 1,000 ml Cat. no. 20012-019 20012-068 20012-043
Quantity 500 ml 10 500 ml (case) 1,000 ml 6 1,000 ml (case) Cat. no. 10010-015 10010-056 10010-031 10010-072

pH: The pH of the 10X solution is 7.1 to 7.3. The pH may change upon dilution to 1X and require readjustment. Intended use: In vitro diagnostic (IVD).

pH: The pH of the 10X solution is 7.3 to 7.5. The pH may change upon dilution to 1X and require readjustment. Intended use: In vitro diagnostic (IVD).
5 Balanced Salt Solutions 141

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Balanced Salt Solutions Preparation

Preparing 1X solutions from10X concentrates
To prepare an acceptable final, single-strength solution, perform the following procedure under aseptic conditions. We recommend Distilled Water (base cat. no. 15230) and 7.5% Sodium Bicarbonate Solution (base cat. no. 25080) for use in this protocol. Use sterile solutions of 1 N sodium hydroxide or hydrochloric acid prepared by membrane filtration using appropriate filters. Also note that, because we adjust the pH of all 10X balanced salt solution concentrates for solubility, you may need to adjust the pH after dilution and add sodium bicarbonate, as appropriate. 1. Aseptically dilute 100 ml of 10X concentrate with approximately 850 ml of sterile distilled water. 2. Aseptically add the correct amount of 7.5% sodium bicarbonate solution. See the table below for concentrations. 3. Adjust the pH as necessary with 1 N hydrochloric acid or 1 N sodium hydroxide. See the table below for 1X pH values. 4. QS to volume with sterile distilled water. 5. Dispense the solution into sterile containers. Cap the bottles tightly with sterile closures and store them at the recommended temperature.
Preparing powdered Dulbeccos PhosphateBuffered Saline (DPBS)

1. To a mixing container that is as close to the final volume as possible, add 23% less distilled water than the desired total volume of DPBS. 2. Add powdered DPBS (less calcium chloride) to room temperature (20C to 30C) water with gentle stirring. Do not heat the water. 3. Rinse the inside of the package to remove all traces of powder. Be sure all material goes into solution before proceeding to the next step. 4. Add 0.1 g of calcium chloride per liter of DPBS. 5. Dilute the DPBS to the desired volume with water and stir until dissolved. Do not overmix. 6. Process the DPBS immediately into sterile containers by membrane filtration using a 0.2 m filter. We recommend using a positivepressure system.
Sodium Bicarbonate Concentrations Achieved Using 7.5% Sodium Bicarbonate Solution (base cat. no. 25080)
NaHCO3 pH of 10X solution Base cat. no. 14065 14080 14185 14200 5.86.1 4.44.7 5.86.1 6.77.0 0.35 0.35 Final concentration (g/L) 7.5% solution to add (ml/L) pH range after 1:10 dilution and addition of NaHCO3 Recommended pH of 1X solution*
Balanced salt solutions 4.7 4.7 7.57.8 5.25.5 6.06.3 7.17.4 7.07.4 7.07.2 7.07.4 7.07.2
* pH range will vary depending on pH of water used for dilution. Formulation does not require addition of NaHCO3.
142
Balanced Salt Solutions Formulations

Dulbeccos Phosphate-Buffered Saline (DPBS)1
Base cat. no. Component Inorganic salts CaCl2 (anhydrous) KCl KH2PO4 MgCl2 (anhydrous) MgCl26H2O NaCl Na2HPO4 Na2HPO47H2O Phenol red D-Glucose Sodium pyruvate 0.10 0.20 0.20 0.10 8.00 2.16 1.00 2.00 2.00 1.00 80.00 21.60 0.20 0.20 8.00 2.16 2.00 2.00 80.00 21.60 0.10 0.20 0.20 0.10 8.00 2.16 1.00 0.036 * 0.20 0.20 0.047 8.00 1.15 0.20 0.20 8.00 1.15 14040 1X liquid, g/L 14080 10X liquid, g/L 14190 1X liquid, g/L 14300 10X liquid, g/L 14287 1X liquid, g/L 21300 Powder, g/L 21600 Powder, g/L
* Supplied separately at 0.1 g/L. 1. J. Exp. Med. (1954) 98, 167.
Hanks Balanced Salt Solutions (HBSS)1

Base cat. no. Component Inorganic salts CaCl2 (anhydrous) KCl KH2PO4 MgCl26H2O* MgSO47H2O NaCl NaHCO3 Na2HPO4 Na2HPO47H2O Other components D-Glucose Phenol red 1.00 10.00 1.00 0.01 1.00 10.00 1.00 0.01 0.14 0.40 0.06 0.10 0.10 8.00 0.35 0.048 1.40 4.00 0.60 1.00 1.00 80.00 0.90 0.40 0.06 8.00 0.35 0.048 0.40 0.06 8.00 0.35 0.048 4.00 0.60 80.00 0.90 0.14 0.40 0.06 0.10 0.10 8.00 0.35 0.048 14025 1X liquid, g/L 14065 10X liquid, g/L 14170 1X liquid, g/L 14175 1X liquid, g/L 14185 10X liquid, g/L 24020 1X liquid, g/L
* MgCl26H2O added to original formulation. Original formulation calls for 0.06 g/L Na2HPO 42H2O. Proc. Soc. Exp. Biol. Med. (1949) 71, 196. Original formulation calls for 0.02 g/L. Proc. Soc. Exp. Biol. Med. (1949) 71, 196. 1. Hanks, J.H., and Wallace, R.E. (1949) Proc. Soc. Exp. Biol. Med. 71, 196, ModificationNational Institutes of Health.
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Phosphate-Buffered Saline (PBS)

Base cat. no. Component Inorganic salts KH2PO4 NaCl Na2HPO47H2O 0.144 9.00 0.795 0.21 9.00 0.726 1.44 90.00 7.95 2.10 90.00 7.26 10010 1X liquid, g/L 20012 1X liquid, g/L 70011 10X liquid, g/L 70013 10X liquid, g/L
Earles Balanced Salt Solutions (EBSS)1

Base cat. no. Component Inorganic salts KCl NaCl NaHCO3 NaH2PO4H2O* Other components D-Glucose 1.00
* Original formulation calls for 150.0 mg/L NaH2PO 42H2O. In Vitro (1974) 9:6.1. J. Nat. Cancer Inst. (1943) 4, 167.
14155 1X liquid, g/L 0.40 6.80 2.20 0.14
144
Notes:
5 Balanced Salt Solutions 145

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Notes:
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Chapter 6 Standard Cell Culture Reagents

Cell Culture Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148 new ACK Lysing Buffer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 Amino Acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 GlutaMAX-I Supplement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 L-Glutamine-200 mM, liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 MEM Amino Acids Solution (50X), liquid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 MEM Non-essential Amino Acids Solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149 Antibiotics and Antimycotics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Actinomycin D . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Ampicillin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Antibiotic-Antimycotic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Blasticidin S HCl . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151 Puromycin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151 Carbenicillin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151 Geneticin Selective Antibiotic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152 Gentamicin Reagent. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153 Hygromycin B . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153 Kanamycin Sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 153 Mycophenolic Acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154 Neomycin Sulfate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154 Penicillin-Streptomycin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154 Penicillin-Streptomycin-Glutamine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154 Penicillin-Streptomycin-Neomycin (PSN) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Polymyxin B Sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Streptomycin Sulfate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155 Zeocin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .155 Antifoaming and Defoaming Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156 Biological Buffers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157 Cell Dissociation Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158 Distilled Water . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161 new WFI for Cell Culture (Water for Injection) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161 Attachment Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162 Growth Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163 Hybridoma Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166 Insect Medium Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167 Mycoplasma Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167 Other Cell Culture Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 168 Cell Culture Reagent Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169 new Presto Blue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 170
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Cell Culture Reagents

Quality Control
For specific information on quality control, please refer to the individual product descriptions. Each production batch is analyzed as required by specifications. Testing may include: Chemical checks. Osmolality and pH are tested to verify that batches meet product specifications. Microbiological tests. Samples of liquid and lyophilized products are tested to confirm the absence of bacterial and fungil contamination using a protocol or the method referenced in the current edition of the U.S. Pharmacopeia sterility test monograph. We also test products of animal origin for the absence of mycoplasmas using the large-volume inoculation method of Barile and Kern 1 and/or the DNA staining method of Chen.2 For details of these tests, see Chapter 5, Animal Sera, Microbiological Tests. Stability testing program. The expiration date shown on every in vitro diagnostic (IVD)-labeled product is an indication of how long the product will deliver effective performance. Our stability testing program establishes and verifies product expiration dates. Product groups are monitored at periodic intervals to determine the length of time that the product will deliver acceptable performance under recommended storage conditions. Functional growth tests. Products are tested for functionality, growth promotion, and cytotoxicity as appropriate. Attention: Researchers using serum-free or reduced-serum conditions or other sensitive cell culture systems should determine cytotoxicity for their specific applications.
Custom Products and Packaging

Many of the dry chemicals used in the manufacture of our products are available in bulk. While they may not be listed in this chapter, please contact our Custom Products Services goup for availability, minimum order, and minimum packagingr. Biochemicals are specified as USP, ACS, FCC, or NF grade as applicable. If no referenced specifications exist, standards are established by Invitrogen. Material Safety Data Sheets or Certificates of Analysis are available upon request. All of these products are intended for use by qualified professionals who will safely handle and dispose of them.
References
1. Barile, M.F. and Kern, J. (1971) Proc. Soc. Exp. Biol. Med. 138, 432. 2. Chen, T.R. (1977) Exp. Cell Res. 104, 255.
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ACK Lysing Buffer

new
L-Glutamine
USP Grade. C5H10N2O3 Formula weight: 146. Recommended storage conditions: 15C to 30C.
ACK Lysing Buffer
ACK Lysing Buffer is suitable for use in the lysis of red blood cells in EDTA-treated whole-blood samples. One volume of ACK Lysing Buffer is sufficient to lyse 1020 times the volume of the blood sample. Recommended storage conditions: 15C to 30C.
Quantity 1 kg Cat. no. 21051-040
Quantity 100 ml Cat. no. A1049201
MEM Amino Acids Solution (50X), liquid

Prepared in distilled water. The amino acids in this solution are 50X the concentration in a standard MEM Medium. For product formulation please refer to the tables at the end of this chapter. pH: 0.6 to 1.2. Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD). Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Amino Acids
GlutaMAX-I Supplement can be used as a direct substitute for L-glutamine for most cell types. GlutaMAX-I Supplement is only removed from the medium by cell metabolism. There is no build-up of toxic metabolites by spontaneous breakdown. 200 mM supplied in 0.85% NaCl. Extremely stable in aqueous solution. See Appendix I, page 202. Recommended storage conditions: 5C to 20C.
Quantity 100 ml 20 x 100ml
Cat. no. 11130-036 11130-077
MEM Non-essential Amino Acids Solution

10 mM (100X), liquid
Reference
1. Christie, A. and Butler, M. (1994) FOCUS 16, 9.
Contains the dipeptide L-alanyl-L-glutamine.
Cat. no. 35050-038 35050-087
Prepared in distilled water. The non-essential amino acids in this solution are 100X the concentration in a MEM- Medium. For product formulation please refer to the tables at the end of this chapter. pH: 0.6 to 1.7. Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD). Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Standard Cell Culture Reagents
L-Glutamine-200 mM, liquid

Supplied at 29.2 mg/ml in 0.85% NaCl. pH: 4.7 to 6.0. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
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Antibiotics and Antimycotics

We supply a wide range of antibiotic mixtures that are useful for the prophylactic control or elimination of a broad spectrum of microbial contaminants from mammalian cell cultures. Some of these antibiotics may also be used in the selection of resistant bacteria. Gibco antibiotic mixtures are subjected to carefully controlled manufacturing procedures to ensure that you always receive products of consistently high quality. Quality control of raw materials. All antibiotics selected for manufacture of gibco antibiotic mixtures are received from approved suppliers. Antibiotic mixtures are formulated according to the product potency as determined by an approved independent testing laboratory. Final product quality control. Our antibiotic mixtures are tested for the absence of bacteria and fungi, utilizing protocols or those referenced in the USP. The pH and osmolality levels must be within established specifications. Use of antibiotics in cell culture systems. The dilutions indicated in the product listing for each antibiotic mixture usually are sufficient to control a broad spectrum of microbial contaminants in a wide range of mammalian cell types. However, prior to using these antibiotic mixtures, we caution users to be aware of the effects that these dilutions could have on specific cell culture systems. Prescreening potent additives is a recognized quality assurance procedure. Periodic discontinuation of antibiotic use is recommended for several reasons: questionable cell culture techniques can be masked, antibiotic-resistant organisms may arise, and cryptic microbial contaminants may be present and be masked by antibiotic use.
Actinomycin D
C62H86N12O16 (nonsterile) Formula weight: 1,255 (anhydrous). Recommended storage conditions: 2C to 8C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Antibiotic-Antimycotic (100X), liquid

Spectrum: Bacteria, fungi, and yeasts. Recommended storage conditions: 5C to 20C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Fungizone is a trademark of E.R. Squibb & Sons, LLC.
Ampicillin, Sodium Salt, lyophilized

200 mg of ampicillin sodium salt per vial. Applications: Selection of resistant bacteria. Recommended storage conditions: 2C to 8C before rehydration; 5C to 20C after rehydration. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Contains 10,000 units of penicillin (base), 10,000 g of streptomycin (base), and 25 g of amphotericin B per ml, utilizing penicillin G (sodium salt), streptomycin sulfate, and amphotericin B as Fungizone Antimycotic in 0.85% saline.
References
1. Miller, J.H. (1972) Experiments in Molecular Genetics, p. 433, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. 2. Davis, R.W., Botstein, D., and Roth, J.R. (1980) Advanced Bacterial Genetics, p. 204, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
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Blasticidin S HCl
C17H26N8O5HCl Formula weight: 458.9. Blasticidin S is a nucleoside selection agent isolated from Streptomyces griseochromogenes. It is a potent translational inhibitor in both prokaryotic and eukaryotic cells.1 E. coli strains are generally sensitive to 50 g/ml while mammalian cells are sensitive to concentrations from 2 to 10 g/ml. Cell death occurs rapidly at low concentrations. Resistance to blasticidin is conferred by the products of the bsd gene from Aspergillus terreus.2 Blasticidin-resistant mammalian cell lines can be generated in as little as seven days post-transfection. Recommended storage conditions: Blasticidin S HCl is provided as a powder and should be stored at 20C. Blasticidin is guaranteed stable for 6 months when properly stored.
Blasticidin and the blasticidin resistance gene (bsd) are the subject of US Patent No. 5,527,701 sold under patent license for research purposes only. Inquiries for commercial use should be directed to: Kaken Pharmaceutical Company, Ltd., Bunkyo Green Court, Center Office Building, 19-20 Fl, 28-8 Honkomagome 2-chome, Bunkyo-ku, Tokyo 113-8650, Japan, Tel: + 81 3-5977-5008; Fax: + 81 3-5977-5008.
Carbenicillin, Disodium Salt

C17H26N8O5HCl Formula weight: 458.9. Application: Used in plant transformation for the killing of Agrobacterium following co-cultivation of the bacteria with plant tissue.1 Recommended storage conditions: 2C to 8C.
Reference
1. Lin, J.J., Assad-Garcia, N., and Kuo, J. (1994) FOCUS 16, 72.
Quantity 5g Cat. no. 10177-012
Quantity 50 mg 20ml 10 x 1ml Cat. no. R210-01 A1113-902 A1113-903
Puromycin
C17H26N8O5HCl C22H29N7O5, 2HCl Formula weight: 458.9. Puromycin Dihydrochloride is an aminonucleoside antibiotic produced by Streptomyces alboniger. It inhibits peptidyl transfer on both prokaryotic and eukaryotic ribosomes. This antibiotic also inhibits the growth of Gram positive bacteria and Morevarious animal and insect cells. Resistance to Puromycin is conferred by the expression of the pac gene. It is widely used in cell biology as selective agent in mammalian and stem cell culture systems. Format: Liquid Solution (Sterile Filtered), 10mgmL Recommended storage conditions: Product shipped in a box containing 10 x 1ml vials for single use, storage -20 deg C.
6 Standard Cell Culture Reagents
Quantity 20ml 10 x 1ml Cat. no. A1113-802 A1113-803
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Geneticin Selective Antibiotic, liquid

50 mg/ml active Geneticin Selective Antibiotic in distilled water. Aseptically filtered. Applications: Geneticin Selective Antibiotic, an aminoglycoside related to gentamicin, 13 is used as a selective antibiotic in molecular genetics experiments.4,5 It is toxic to bacteria,1,2 yeast,3,4 higher plant 5 and mammalian cells,68 and protozoans and helminths.9,10 The resistance genes are dominant and are located on both transposons Tn601(903) and Tn5. The gene on Tn5 is referred to as aminoglycoside phosphotransferase 3(II) or APH(3)II, whereas the gene on Tn601 is referred to as aminoglycoside phosphotransferase 3(I) or APH(3)I. The genes are bacterial in derivation, but they can be expressed in eukaryotic cells. Introduction of either of these genes into cells can confer resistance to Geneticin Selective Antibiotic, which enables the cells to grow in media containing Geneticin Selective Antibiotic. Properties: Specific rotation: 104C to 121C on dried basis in water Formula: C20H40O10N42H2SO4 Molecular weight: 496.5 (base), 692.7 (sulfate) Potency: 700 g/mg Elemental analysis: C = 34.68, N = 8.09, H = 6.40 (sulfate) Heavy metals: <20 ppm as Pb Stability: Stable up to 24 months at 20C or at 2C to 8C. Notes: The amount of Geneticin Selective Antibiotic required in culture media to select for resistant cells varies. For a multiplying cell, 3 to 7 days will be required for death to occur. See the table for guidelines. For a discussion of performance parameters, such as potency and working range, please visit www.invitrogen.com/geneticin. Recommended storage conditions: 20C, or at 2C to 8C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
CH3 HO CH O OH HO NH2 O O OH 2H2SO4 NH2 NH2
HO H3C
O NHCH3 OH
Geneticin Selective Antibiotic.
Geneticin Selective Antibiotic: Guide to Concentration

Cells Dictyostelium Mammalian Plant Yeast
*Active drug
Transposon Tn5 Tn5 Tn5 Tn601
Gene APH(3)-II APH(3)-II APH(3)-II APH(3)-I
Concentration (mg/L) 10* 35* 400* 200* 10* 500 125
Notes Cells grown in medium Cells plated on lyophilized bacteria For selection For maintenance For selection For selection For maintenance
References 8 6, 7 5 3
References
1. Weinstein, M.J., Wagman, G.H., Testa, R.T., and Marquez, J.A. (May 25, 1976) Antibiotic G-418 and the Production Thereof, US Patent 3,959,254. 2. Weinstein, M.J., Wagman, G.H., Testa, R.T., and Marquez, J.A. (December 14, 1976) Antibiotic G-418 and the Production Thereof, US Patent 3,997,403. 3. Jimnez, A. and Davies, J. (1980) Nature 287, 869. 4. Davies, J. and Jimnez, A. (1980) Am. J. Trop. Med. Hyg. 29(5) Suppl: 1089. 5. Ursic, D., Kemp, J.D., and Helgeson, J.P. (1981) Biochem. Biophys. Res. Comm. 101(3), 1031. 6. Southern, P.J. (1981) In Vitro 18, 315. 7. Southern, P.J. and Berg, P. (1982) J. Mol. Appl. Genet. 1, 327. 8. Canaani, D. and Berg, P. (1982) Proc. Natl. Acad. Sci. USA 79, 5166. 9. Loebenberg, D., Counelis, M., and Waitz, J.A. (1975) Antimicrob. Agents Chemother. 7:6, 811. 10. Waitz, J.A., Sabatelli, F., Menzel, F., and Moss, E.L., Jr. (1974) Antimicrob. Agents Chemother. 6:5, 579.
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Geneticin Selective Antibiotic, powder

Crystalline, nonsterile (G418 sulfate) Stability: Very stable, may be stored in powdered form at room temperature (15C to 30C) for up to 3 years; soluble in water. Recommended storage conditions: 15C to 30C before reconstitution; 2C to 8C after reconstitution. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Hygromycin B, liquid
C20H37N3O13 Purity: >80%. Formula weight: 527.5. Assay: 50 mg/ml. Application: Used in plant transformation 1,2 and mammalian transfection systems 3-5 for the selection of resistant cells. Recommended storage conditions: 2C to 8C.
Quantity 1g 5g 10g 25 g Cat. no. 11811-023 11811-031 11811-064 11811-098
References
1. Lin, J.J., Ma, J., Assad-Garcia, N., and Kuo, J. (1996) FOCUS 18, 47. 2. Datta, S.K., Peterhans, A., Datta, K., and Potrykus, I. (1990) Biotech. 8, 736. 3. Gonzales, A., Jimnez, A., Vazquez, D., Davies, J.E., and Schindler, D. (1978) Biochem. Biophys. Acta 521, 459. 4. Blochlinger, K. and Diggelmann, H. (1984) Mol. Cell Biol. 4, 2929. 5. Giordano, T.J. and McAllister, W.T. (1990) Gene 88, 285.
Gentamicin Reagent Solution, liquid

Spectrum: Gram-positive and gram-negative bacteria. Recommended concentration: 0.5 to 50 g/ml. Recommended storage conditions: 15C to 30C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Kanamycin Sulfate
USP grade C18H36N4O11H2SO4 (nonsterile) Molecular weight: 583 Assay: 750 g kanamycin/mg (anhydrous) Recommended storage conditions: 15C to 30C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Size 10 mg 50 mg Quantity 100 ml 20 10 ml (case) 100 ml 20 10 ml (case) Cat. no. 15710-049 15710-080 15750-045 15750-102
Contains 50 mg/ml gentamicin sulfate in distilled water.
Quantity 5g 25 g Cat. no. 11815-024 11815-032
Kanamycin Sulfate (100X), liquid

Spectrum: Gram-positive and gram-negative bacteria. Recommended storage conditions: 5C to 20C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Quantity 100 ml 20 x 100 ml Cat. no. 15160-047 15160-070
Contains 10 mg/ml kanamycin (base) utilizing kanamycin sulfate in 0.85% saline.
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Mycophenolic Acid
Crystalline, nonsterile Applications: Mycophenolic Acid (MPA) is an antibiotic that blocks inosine monophosphate dehydrogenase, an enzyme in the guanosine monophosphate (GMP) synthesis pathway. This product can be used as a selective agent in mammalian systems because of the different effects of the activities of mammalian HPRT (hypoxanthine phosphoribosyl transferase) and bacterial XGPRT (xanthine guanine phosphoribosyltransferase). Both enzymes mediate an alternative (salvage) pathway for GMP synthesis. Because mammalian cells can use xanthine to a limited extent during GMP synthesis, GMP synthesis is effectively blocked by MPA in guanine-free media. Addition of Aminopterin to the medium has been found to enhance this blockage. XGPRT of E. coli can effectively convert xanthine to xanthine monophosphate. The insertion of the Ecogpt gene, which codes for XGPRT, into mammalian cells allows these cells to grow in media containing 25 g/ml of MPA. For more information, see the references below. Properties: Formula: C17H20O6 Molecular weight: 320.35 Purity: 95% Solubility: moderately soluble in ether and chloroform, freely soluble in alcohol, insoluble in water Recommended storage conditions: 15C to 30C before reconstitution; 5C to 20C after reconstitution. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Penicillin-Streptomycin, liquid
Spectrum: Gram-positive and gram-negative bacteria. Recommended concentration: 50100 U of penicillin, 50100 g of streptomycin. Recommended storage conditions: 5C to 20C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Contains 5,000 units of penicillin (base) and 5,000 g of streptomycin (base) per ml utilizing penicillin G (sodium salt) and streptomycin sulfate in 0.85% saline.
Cat. no. 15140-148 15140-122 15140-130
Contains 10,000 units of penicillin (base) and 10,000 g of streptomycin (base) per ml utilizing penicillin G (sodium salt) and streptomycin sulfate in 0.85% saline.
Penicillin-Streptomycin-Glutamine (100X), liquid

Utilizes penicillin G (sodium salt) and streptomycin sulfate. Spectrum: Gram-positive and gram-negative bacteria. Recommended storage conditions: 5C to 20C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
References
1. Cass, C.E., Lowe, J.K., Manchak, J.M., and Henderson, J.F. (1977) Cancer Res. 37, 3314. 2. Lowe, J.K., Brox, L., and Henderson, J.F. (1977) Cancer Res.736. 3. Mulligan, R.C. and Berg, P. (1981) Proc. Natl. Acad. Sci. USA 78, 2072. 4. Gorman, C., Padmanabhan, R., and Howard, B.H. (1983) Science 221, 551.
Contains 10,000 units of penicillin (base), 10,000 g of streptomycin (base), and 29.2 mg of L-glutamine per ml in 0.85% saline, in a 10 mM citrate buffer to maintain penicillin potency.
Neomycin Sulfate
USP grade, nonsterile Toxicity data: ROTECS QP4375000. Recommended storage conditions: 15C to 30C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
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Penicillin-Streptomycin-Neomycin (PSN) Antibiotic Mixture (100X), liquid

Spectrum: Gram-positive and gram-negative bacteria. Recommended storage conditions: 5C to 20C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Zeocin
C55H83N19O21S2Cu Formula weight: 1137.41. Zeocin is an antibiotic in the bleomycin family. It causes cell death by intercalating into the DNA and cleaving it. Resistance to Zeocin is conferred by the Sh ble gene, which encodes a 13,665 DA protein. This protein confers resistance to Zeocin by binding to the antibiotic and preventing it from binding DNA. Zeocin is effective on most aerobic cells and can be used for selection in mammalian cell lines, yeast, and bacteria. The amount of Zeocin required for selection will vary with each cell type. Recommended storage conditions: Zeocin is provided in solution at a concentration of 100 mg/ml. Store Zeocin at 20C in the dark. The drug is guaranteed stable for 6 months when properly stored.
Zeocin is a trademark of CAYLA. The ble resistance gene is the subject of one or more of US Patent Nos. 5,021,344 and 5,118,620 and foreign equivalents owned by Cayla and licensed to Invitrogen. This product is sold for research purposes only. For commercial license information, please contact: Licensing Department, Invitrogen Corporation, 5791 Van Allen Way, Carlsbad, CA 92008. Phone: 760-603-7200, Fax: 760-602-6500.
Contains 5 mg of penicillin, 5 mg of streptomycin, and 10 mg of neomycin per ml utilizing penicillin G (sodium salt), streptomycin sulfate, and neomycin sulfate in 0.85% saline.
Polymyxin B Sulfate
USP grade, nonsterile, crystalline Spectrum: Gram-negative bacteria. Recommended storage conditions: 15C to 30C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Quantity 25 MU Cat. no. 21850-029
Quantity 1g 5g Cat. no. R250-01 R250-05
Streptomycin Sulfate
USP grade (C21H39N7O12)23H2SO4 (nonsterile) Formula weight: 1,457. Assay: 650 to 850 g/mg (anhydrous). Application: Selection of resistant bacteria. Spectrum: Gram-negative bacteria. Recommended storage conditions: 2C to 8C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling
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Antifoaming and Defoaming Reagents

FoamAway Irradiated
Antifoaming/defoaming reagent. Packaged in a convenient and versatile media bag that can be coupled to your bioreactor via a sterile connection. Can be used with both mammalian and microbial cell culture systems. FoamAway Irradiated is specifically designed to offer high performance and convenience in combating foam in your cell culture system. It is prediluted, presterilized, and packaged to connect directly to your bioreactor with no further processing required. FoamAway Irradiated consists of a 30% simethicone emulsion diluted with WFI (water for injection)-grade purified water to a final concentration of 3%. This material is then packaged into flexible media bags and irradiated. Gamma-irradiation sterilization of the FoamAway product family has been validated to a sterility assurance level (SAL) of 10-6. The radiation validation study was based on the Association for the Advancement of Medical Instrumentation (AAMI) Technical Information Report (TIR) No. 33:2005 standard, entitled Sterilization of Health Care Products RadiationSubstantiation of a Selected Sterilization DoseMethod VDmax. This process validates a sterilization dose based on the determination of the product bioburden and comparison of that bioburden information to a model population having a defined resistance to radiation. AAMI TIR No. 33:2005 was designed to meet the requirements of ANSI/AAMI/ISO 11137, entitled Sterilization of Health Care ProductsRadiation Sterilization. Please note that FoamAway Irradiated cannot be filtered due to the size of the simethicone particles. Recommended storage conditions: room temperature; shake well before use; do not filter.
FoamAway Irradiated AOF
Quantity 500 ml in 1 L bag 2.5 L in 5 L bag Cat. no. 006-0096BC 006-0096BA
Ready-to-use, animal originfree reagent for both antifoaming and defoaming applications Can be used with both mammalian and microbial cell culture systems Formulation and media bag contact layer contain no components derived from human or animal sources FoamAway Irradiated AOF is specifically designed to offer high performance and convenience in combating foam in your cell culture system. It is prediluted, presterilized, and packaged to connect directly to your bioreactor with no further processing required. FoamAway Irradiated AOF consists of an animal originfree 30% simethicone emulsion diluted with WFI (water for injection)-grade purified water to a concentration of 3%. This material is then packaged into flexible media bags and irradiated. Gamma-irradiation sterilization of the FoamAway product family has been validated to a sterility assurance level (SAL) of 106. The radiation validation study was based on the Association for the Advancement of Medical Instrumentation (AAMI) Technical Information Report ( TIR) No. 33:2005 standard, entitled Sterilization of Health Care ProductsRadiationSubstantiation of a Selected Sterilization DoseMethod VDmax. This process validates a sterilization dose based on the determination of the product bioburden and comparison of that bioburden information to a model population having a defined resistance to radiation. AAMI TIR No. 33:2005 was designed to meet the requirements of ANSI/AAMI/ISO 11137, entitled Sterilization of Health Care ProductsRadiation Sterilization. Please note that FoamAway Irradiated AOF cannot be filtered, due to the size of the simethicone particles. Recommended storage conditions: room temperature; shake well before use; do not filter. For more information, visit www.invitrogen.com/foamaway.
Quantity 500 ml in 1 L bag 2.5 L in 5 L bag Cat. no. A1036902 A1036901
For more information about additional components exclusively available through Invitrogen, such as an alternative for recombinant human insulin, please contact our Custom Products Services group or www.invitrogen.com/limitededition.
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Biological Buffers
The growth of mammalian cells in culture media depends on both the pH of the media and the species of buffer present. Also, a high buffer capacity with a low buffer concentration is needed to avoid an undesirable fluctuation of pH due to tissue metabolism. Fluctuation of pH may lead to inhibition of growth or have other undesirable effects on cell parameters. Disadvantages and limitations of bicarbonate/CO2 and phosphate buffers are well known, and Tris buffers are cytotoxic in certain cases. HEPES is the most generally used zwitterionic buffer. Improved pH control between pH 6.7 and 8.4 may be obtained when 10 to 25 mM HEPES is incorporated into culture media. However, as buffer species vary in their toxicity to different types of cells, care must be taken in choosing buffers and their concentrations for use with new cell lines.
HEPES Buffer Solution (1 M)

pH: 7.2 to 7.5. pKa: 7.3 at 37C. Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD).
Sodium Bicarbonate Solution, 7.5%

7.5 g/100 ml Recommended storage conditions: 15C to 30C.
Quantity 100 ml Cat. no. 15630-049 15630-056 15630-122 Cat. no. 25080-060
Contains 238.3 g/L prepared in distilled water.
HEPES Ultrapure
(N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid) C8H18N2O4S Formula weight: 238. Purity: >99.5%. Useful pH range: 6.8 to 8.2. Recommended storage conditions: 15C to 30C
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Cell Dissociation Products

Cell Dissociation Buffers
Cell Dissociation Buffer, enzyme free, Hanks-based Cell Dissociation Buffer, enzyme free, PBS-based
Cell dissociation buffers are membrane-filtered, isotonic, and enzyme-free aqueous formulations of salts, chelating agents, and cell-conditioning agents in either Ca2+- and Mg2+-free Hanks balanced salt solution or Ca2+- and Mg2+-free phosphate-buffered saline. They are suitable for gentle dissociation of mammalian cells from support substrates and each other, and for studies requiring intact cell-surface proteins such as ligand binding, flow cytometry, and immunohistochemistry.1,2 These buffers are not recommended for routine passage of strongly adherent cell types. Performance and quality testing: Buffers are membrane filtered and mycoplasma tested. Cell integrity after dissociation with buffers is measured by a plating efficiency assay and a serotonin receptor function assay. Recommended storage conditions: 15C to 30C.
Collagenase Type II
Prepared from Clostridium histolyticum for dissociation of tissues. Suggested for preparation of cells from liver, bone, thyroid, heart, and salivary gland tissue. Recommended storage conditions: 2C to 8C.
Collagenase Type IV
Prepared from Clostridium histolyticum for dissociation of tissues from pancreas. For isolation of pancreatic islets. Recommended storage conditions: 2C to 8C.
References
1. Julius, D., Livelli, T.J., Jessell, T.M., and Axel, R. (1989) Science 244, 1057. 2. Livelli, T.J. (1989) FOCUS 11, 74.
Quantity 100 ml (Hanks-based) 100 ml (PBS-based) Cat. no. 13150-016 13151-014
Dispase
Prepared from Bacillus polymyxa. For gentle dissociation of animal tissues to release individual cells. Recommended storage conditions: 2C to 8C.
Collagenase, lyophilized
(Clostridiopeptidase A). Prepared from Clostridium histolyticum. E.C. number: 3.4.2.4.3 Activity: 125 units/mg weight. One unit will liberate 1 mol amino acid from collagen, expressed as L-leucine, in 18 hr at 37C. Recommended storage conditions: 2C to 8C.
Collagenase Type I
Prepared from Clostridium histolyticum for dissociation of tissues. Suggested for epithelial, lung, fat, and adrenal tissue cell preparations. Recommended storage conditions: 2C to 8C.
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Trypsin Solutions
Gibco trypsin solutions are used widely for dissociation of tissues and cell monolayers. We can provide cell culture dissociating solutions that will perform consistently, batch after batch. The biochemical assays performed on Trypsin (1:250) determine both the trypsin-specific activity and the level of certain copurified enzymes that influence cell removal and viability. All trypsin solutions are prepared from porcine parvovirustested and mycoplasma-tested materials. Gibco trypsin solutions are prepared under controlled conditions, using accelerated manufacturing procedures to minimize loss of enzyme activity from denaturation and autodegradation. Raw material quality control. Trypsin powdered raw material is subjected to standard assays for solubility, loss of weight on drying, residue on ignition, and trypsin activity as outlined in the current edition of the U.S. Pharmacopeia, National Formulary (USP/NF).
Powdered Trypsin
We offer trypsin preparations as nonsterile powders.
Gamma-Irradiated Liquid and Powdered Trypsin

Upon request, trypsin can be gamma-irradiated. We have validated the use of gamma-irradiation to inactivate certain porcine viruses which may be present in trypsin. Our studies indicate that an irradiation dose of 25 kGy will inactivate virus while preserving the biological performance in cell culture and the trypsin enzymatic activity. Additionally, osmolality and pH remain unchanged. For further information about our validation results, contact the Invitrogen Technical Support.
Trypsin Replacement Enzymes

TrypLE trypsin replacement enzyme is a microbially produced alternative to animal trypsin. It is manufactured in a controlled fermentation process that is completely free of animal- and human-derived components. The process produces an exceptionally pure reagent that is gentle on cells. Inactivation with trypsin inhibitors is not required. GIBCO TrypLE enzyme can be stored refrigerated and is stable at room temperature for 6 months. Visit www.invitrogen.com/trypleexpress for more information.
Accepted values
Activity: One USP trypsin unit is the activity causing a change in absorbance of 0.003/min in the presence of N-benzoyl-L-arginine ethyl ester hydrochloride at 25C, pH 7.6. Trypsin (1:250) has an activity such that 1 g will digest (hydrolyze) 250 g of casein under the conditions of the USP test for protease activity of pancreatin. Porcine parvovirus: None detected. Mycoplasma: None detected. Amylase activity: Acceptable. Nucleotidase activity: Acceptable. Final product quality control: GIBCO enzyme preparations are tested to confirm the absence of bacterial and fungal contamination, employing protocols referenced in the current USP. The pH and osmolality of all batches are monitored using calibrated measuring equipment to verify conformance with product specifications. In vitro bioassays are performed on the final product to verify the ability of the trypsin solutions to remove an adherent cell line from the substratum while preserving cellular integrity. The cell line selected for this assay is a human transformed cell line. The trypsin-dependent cell-removal times must be within a range established for each cell line. Following trypsinization, cells are replated and morphology is monitored after 18 to 24 hr. Trypsin removal and replating are tested in parallel with a reference trypsin solution. For more information about GIBCO enzyme preparations, contact Invitrogen Technical Support.
TrypLE Select Stable Trypsin Replacement Enzyme (1X), liquid
An animal originfree cell dissociation enzyme. This recombinant enzyme is an alternative to animal trypsin for the dissociation of adherent mammalian cells from plasticware. TrypLE Select is manufactured for the industrial market and has release testing for both endotoxin and enzyme activity. This material also has a Drug Master File on record. Formulated in D-PBS with 1 mM EDTA. Stable at 15C to 30C for 6 months. Recommended storage conditions: 2 to 8C.
Contains no phenol red.
Cat. no. 12563-011 12563-029
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TrypLE Express, Stable Trypsin Replacement Enzyme (1X), liquid
Trypsin-EDTA (0.05% Trypsin, EDTA 4Na) (1X), liquid

Porcine parvovirus and mycoplasma tested. Recommended storage conditions: 5C to 20C.
A pure enzyme for gentler cell dissociation. This recombinant enzyme is an alternative to animal trypsin for the dissociation of adherent mammalian cells from plasticware. TrypLE Express is animal originfree and thus ideal for both serum and serum-free cultures. Formulated in D-PBS with 1 mM EDTA. Stable at 15C to 30C for 6 months. Recommended storage conditions: 2C to 8C.
Quantity 100 ml 10 500 ml (case) 20 100 ml (case) 500 ml Cat. no. 25300-054 25300-104 25300-096 25300-062
Quantity 100 ml 500 ml 20 100 ml (case) 5L
Contains phenol red.
Cat. no. 12605-010 12605-028 12605-036 12605-093
Contains phenol red, 0.5 g/L of trypsin (1:250) and 0.2 g/L of EDTA4Na in Hanks Balanced Salt Solution without CaCl2, MgCl26H2O, or MgSO47H2O.
Trypsin-EDTA (0.5% Trypsin, EDTA4Na) (10 X), liquid

Porcine parvovirus and mycoplasma tested. Dilution: Aseptically prepare 1X solution in balanced salt solution without Ca++ or Mg++. Final concentration: 0.5 g/L of trypsin and 0.2 g/L of EDTA4Na (1:250). Recommended storage conditions: 5C to 20C.
Quantity 100 ml 500 ml 5L 20 100 ml (case)

Cat. no. 12604-013 12604-021 12604-054 12604-039
Contains 5.0 g/L of trypsin (1:250), 2.0 g/L of EDTA4Na, and 8.5 g/L of NaCl but no phenol red.
Trypsin, 0.25% (1X), liquid

Porcine parvovirus and mycoplasma tested. Source: Porcine. Recommended storage conditions: 5C to 20C.
Trypsin-EDTA (0.25% Trypsin, EDTA4Na) (1X), liquid

Cat. no. 25050-014 25050-030
Porcine parvovirus and mycoplasma tested. Recommended storage conditions: 5C to 20C.
Quantity 100 ml 20 100 ml (case) 500 ml Cat. no. 25200-056 25200-114 25200-072
Contains phenol red, 2.5 g/L of trypsin (1:250) in Hanks Balanced Salt Solution without CaCl2, MgCl26H2O, or MgSO47H2O.
Trypsin, 2.5% (10X), liquid

Porcine parvovirus and mycoplasma tested. Source: Porcine. Dilution: Aseptically prepare 1X solution in balanced salt solution without Ca++ or Mg++. Final concentration: 2.5 g/L of trypsin (1:250). Recommended storage conditions: 5C to 20C.
Contains phenol red, 2.5 g/L of trypsin (1:250), and 0.38 g/L of EDTA4Na in Hanks Balanced Salt Solution without CaCl2, MgCl26H2O, or MgSO47H2O.
Contains 25 g/L of trypsin (1:250) and 8.5 g/L of NaCl but no phenol red.
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Trypsin (1:250), powder

Porcine parvovirus and mycoplasma tested. Activity: 1 g of trypsin will digest 250 g of casein substrate under the conditions of the USP monographs for protease activity of pancreatin (25C for 10 min at pH 7.6).1 Recommended storage conditions: 2C to 8C.
Distilled Water
Water, distilled
Membrane filtered and endotoxin screened. Recommended storage conditions: 15C to 30C.
Reference
1. United States Pharmacopeia.
Quantity 100 ml Cat. no. 27250-018 20 100 ml (case) 500 ml 10 500 ml (case) 1L 6 1000 ml 5L 10 L Cat. no. 15230-071 15230-196 15230-089 15230-204 15230-097 15230-001 15230-253 15230-261
Quantity 100 g
Trypsin Inhibitor, soybean

Activity: >7,000 BAEE units of inhibition per mg. Recommended use: 0.25 to 0.5 mg/ml in PBS to inactivate an equal volume of trypsin. Recommended storage conditions: 5C to 20C.
WFI for Cell Culture (Water for Injection)

new
Versene 1: 5,000 (1X), liquid

Concentration: 1X = 0.2 g/L = 0.48 mM. Recommended storage conditions: 2C to 8C.
Pure Always
gibco WFI for Cell Culture is high quality, cell culture grade water, research use only. Every batch of gibco WFI for Cell Culture is membrane-filtered to 0.1 m and undergoes strict quality-control testing for sterility, endotoxin levels, conductivity, osmolality, oxidizable substances, and pH. gibco WFI for Cell Culture meets the USP monograph for water for injection Recommended storage conditions: gibco WFI for Cell Culture, USP water for injection, research use only has a 12 month shelf life at 15-30C.
Quantity 100 ml
Contains 0.2 g/L EDTA4Na in phosphate-buffered saline.
Cat. no. 15040-033
Quantity 500ml 1000ml 10X500ml 6x1L Cat. no. A1287-301 A1287-302 A1287-303 A1287-304
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Attachment Factors
Fibronectin, Human Plasma
Fibronectin is a large cell-surface and plasma protein that exhibits structural and adhesive properties in cell-associated fibrillar matrices. Fibronectin is one of the primary cell adhesion molecules. Human plasma fibronectin is isolated from human plasma by affinity chromatography. Fibroncetin has a molecular weight of 440,000 and is made up of two 220,000 DA polypeptides. Human fibronectin is filtered (0.2 m) and supplied lyophilized. Performance and quality testing: Purity is determined by SDS-PAGE. Human fibronectin is assayed for its ability to support cell attachment and spreading, and screened to confirm absence of mycoplasmas or bacterial contamination. Recommended storage conditions: 5C to 20C. Notes: Do not store in dilute solution. Avoid repeated freezing and thawing. Caution: Human-origin materials are nonreactive (donor level) for anti-HIV 1 and 2, anti-HCV, and HBs Ag. Handle in accordance with established biosafety practices.
Natural Mouse Laminin

Natural Mouse Laminin is purified from the Engelbreth-Holm-Swarm (EHS) sarcoma and is provided in Tris-buffered saline. Performance and quality testing: Purity is determined by SDS-PAGE. Mycoplasma content was determined to be negative. Functional performance is verified by cell attachment. Recommended storage conditions: 20C. Notes: Do not store in dilute solution. Avoid repeated freezing and thawing.
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Growth Supplements
250X Cholesterol Lipid Concentrate is a chemically defined, protein-free supplement used for growing NS0 and other lipid dependent cell lines to high densities while supporting growth and protein expression without complicating downstream purification. The supplement can be added to 1X media such as CD Hybridoma Medium and filtered with minimal loss of lipids. Custom formulations and packaging configurations as well as supplements used in feeding strategies are available through our Custom Products Services group. Recommended storage conditions: 2C to 8C, in the dark.
AlbuMAX I
Lipid-rich bovine serum albumin for cell culture.1 It is chromatographically purified and has an IgG content 0.1%. Recommended storage conditions: 2C to 8C.
Quantity 25 g 1 kg Cat. no. 11020-021 11020-039
Reference
1. Gorfien, S., Paul, W., Walowitz, J., Keem, R., Biddle, W., and Jayme, D., Growth of NS0 cells in protein-free, chemically defined medium (2000) Biotechnol. Prog. 16: 682-687.
AlbuMAX II
Lipid-rich bovine serum albumin for cell culture.1 It is chromatographically purified and has an IgG content <1.0%. Recommended storage conditions: 2C to 8C.
Reference
1. Airey, P., Benny, G., Swartzwelder, F., and Conrad, D. (1991) FOCUS 13, 2.
Related products Page Hybridoma products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 106
Quantity 25 g 100 g 1 kg Cat. no. 11021-029 11021-037 11021-045

Chemically Defined Lipid Concentrate is a concentrated lipid emulsion designed to reduce or replace fetal bovine serum in cell culture media for a wide variety of applications, including growth and maintenance of CHO, hybridoma, and insect cells in culture; monoclonal antibody production by hybridomas; and viral expression in insect cells. The components of this medium supplement are chemically defined. For product formulation please refer to the tables at the end of this chapter. Insect cell culture tested. Recommended storage conditions: 2C to 8C, in the dark. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Bovine Albumin Fraction V Solution (7.5%)
7.5 g/100 ml Prepared with Bovine Serum Albumin Fraction V in phosphate-buffered saline. Recommended storage conditions: 2C to 8C.
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B-27 Serum-free Supplements

B-27 Serum-free Supplements are 50X liquid, serum-free supplements for growth and long-term viability of hippocampal neurons. 1,2 These supplements are used in combination with a basal medium such as Neurobasal Medium.2 B-27 Supplement Minus AO is the complete B-27 Supplement without 5 antioxidants (vitamin E, vitamin E acetate, superoxide dismutase, catalase, and glutathione) that would interfere with studying free-radical damage to neurons. B-27 Supplement Minus Vitamin A is the complete B-27 Supplement without vitamin A. The omission of vitamin A in this supplement prevents neural differentiation of stem cells. Recommended storage conditions: 5C to 20C.
Insulin-Transferrin-Selenium Supplements (ITS)

Gibco Insulin-Transferrin-Selenium supplements reduce the fetal bovine serum (FBS) concentration required for routine maintenance and low-density attachment of many cell types. Two supplements are available for use with noncomplex media such as RPMI 1640 or Earles Minimum Essential Medium: Insulin-Transferrin-Selenium-G for general use and Insulin-Transferrin-Selenium-A for adherent cell cultures. In addition, a third supplement, Insulin-TransferrinSelenium-X, is available for use with complex media that contain sodium pyruvate, such as F-12 Nutrient Mixture, Dulbeccos Minimum Essential Medium/F-12 Nutrient Mixture, or Earles Minimum Essential Medium. All of these supplements are supplied at 100X and should be added to media at a ratio of 10 ml per liter of medium. Initially, FBS must be added at a concentration of 2% to 4% to achieve growth comparable to the same medium supplemented with 10% FBS; FBS concentration can be reduced further following initial adaptation. Performance testing: Not tested to determine plating efficiency.
References
1. Brewer, G.J., Torricelli, J.R., Evege, E.K., and Price, P.J. (1993) J. Neuroscience Res. 35, 567. 2. Brewer, G.J., Torricelli, J.R., Evege, E.K., and Price, P.J. (1994) FOCUS 16, 6.
Product B-27 Serum-free Supplement (50X), liquid B-27 Supplement Minus AO (50X), liquid B-27 Supplement Minus Vitamin A (50X), liquid Quantity 10 ml 10 ml 10 ml Cat. no. 17504-044 10889-038 12587-010
Insulin-Transferrin-Selenium-G Supplement (100X)

Prepared in Earles Balanced Salt Solution (EBSS). For use in general culture applications. Recommended storage conditions: 2C to 8C.
Quantity 10 ml
Related products Page Neurobiology products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .114
Cat. no. 41400-045
G-5 Supplement (100X), liquid

G-5 Supplement is a serum-free supplement for the growth and expression of glial cells (normal and tumor) of astrocytic phenotype. For product formulation please refer to the tables at the end of this chapter. Recommended storage conditions: 5C to 20C.
Insulin-Transferrin-Selenium-A Supplement (100X)

Prepared in EBSS. For use in adherent (anchorage-dependent) cultures. Recommended storage conditions: 2C to 8C.
Quantity 10 ml
Cat. no. 51300-044
Insulin, Human Recombinant, Zinc

4 mg/ml Potency: (dry basis) 27 USP units/mg. Suggested range for supplementation: 2 to 20 g/ml. Recommended storage conditions: 5C to 20C.
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Insulin-Transferrin-Selenium-X Supplement (100X)

Prepared in EBSS. For use with complex media that contain sodium pyruvate. Recommended storage conditions: 2C to 8C.
Transferrin, Bovine (APO form), lyophilized

Iron <50 g/g protein. Suggested range for supplementation: 0.5 to 50 g/ml. Recommended storage conditions: 2C to 8C.
Quantity 10 ml
Contains no phenol red or sodium pyruvate.
Cat. no. 51500-056 Quantity 1g Cat. no. 11108-016
Transferrin, Bovine (holo form), lyophilized Insulin-Transferrin-Selenium Supplement Formulations

Supplement ITS-G ITS-A ITS-X Insulin (1.0 g/L) Sodium selenite (0.67 mg/L) Transferrin (0.55 g/L) Sodium pyruvate (11.0 g/L) Ethanolamine (0.2 g/L)
90% iron saturated. Suggested range for supplementation: 0.5 to 50 g/ml. Recommended storage conditions: 2C to 8C.
Quantity 100 mg 25 g Cat. no. 11107-018 11107-047
Notes: Supplements have been prepared in EBSS without phenol red. All formulations are 100X.
Lactalbumin Hydrolysate, powder (extra soluble)

Tissue-culture grade. Recommended storage conditions: 15C to 30C.
Transferrin, Bovine (partially iron loaded), lyophilized

Iron content 400 to 500 g/g protein. Suggested range for supplementation: 0.5 to 50 g/ml. Recommended storage conditions: 2C to 8C.

N-2 Supplement is a serum-free supplement for the growth and expression of post-mitotic neurons and tumor cells of neuronal phenotype. For product formulation please refer to the tables at the end of this chapter. Recommended storage conditions: 5C to 20C.
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Hybridoma Reagents
We offer high-quality reagents required for selecting hybridomas. Selection and confirmation of hypoxanthine guanine phosphoribosyl transferase (HGPRT )deficient cells is achieved by use of the toxic base analog 8-azaguanine. HGPRT mutants will continue to proliferate, whereas cells that possess HGPRT will die. HAT medium (medium supplemented with hypoxanthine, aminopterin, and thymidine), which is used post-fusion as a selective medium, serves a dual purpose. Aminopterin, as an inhibitor of de novo nucleoside biosynthesis, is used to select against unfused myeloma cells and myeloma-myeloma fused partners. Hypoxanthine and thymidine supply preformed purines and pyrimidines for DNA synthesis by hybridomas via salvage pathways, as a result of the contribution of HGPRT by spleen cells. HT medium (medium supplemented with hypoxanthine and thymidine) is used as a rescue medium that continues providing preformed purines and pyrimidines downstream to overcome the effects of residual intracellular aminopterin. Related products Page Hybridoma products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .106

HAT Supplement (50X), liquid

HAT Supplement (50X) is a liquid mixture of sodium hypoxanthine, aminopterin, and thymidine. After dilution, HAT is suitable for use as a post-fusion selective medium to eliminate unfused or self-fused HGPRT-myeloma cells. Hypoxanthine and thymidine supply preformed purines and pyrimidines for DNA synthesis by hybridomas via the salvage pathway that utilizes HGPRT contributed by the fused spleen cell. Aminopterin, a folic acid antagonist, inhibits the de novo nucleoside biosynthesis pathway. Concentration at 50X: 5 mM sodium hypoxanthine, 20 M aminopterin, 0.8 mM thymidine. Recommended storage conditions: 5C to 20C.

250X Cholesterol Lipid Concentrate is a chemically defined, protein-free supplement used for growing NS0 and other lipid-dependent cell lines to high densities while supporting growth and protein expression without complicating downstream purification. The supplement can be added to 1X media such as CD Hybridoma and filtered with minimal loss of lipids. Custom formulations and packaging configurations as well as supplements used in feeding strategies are available through our Custom Products Services group. Recommended storage conditions: 2C to 8C, in the dark.
Reference
1. Gorfien, S., Paul, W., Walowitz, J., Keem, R., Biddle, W. and Jayme, D., Growth of NS0 cells in protein-free, chemically defined medium (2000) Biotechnol. Prog. 16: 682687.
HT Supplement (100X), liquid

HT Supplement (100X) liquid is a mixture of sodium hypoxanthine and thymidine. After dilution, HT Supplement is suitable for use as a rescue medium. The supplement provides preformed purines and pyrimidines to overcome the effects of residual intracellular aminopterin. Once the de novo biosynthesis pathway for nucleosides has been reestablished, HT supplementation can be discontinued. Concentration at 100X: 10 mM sodium hypoxanthine, 1.6 mM thymidine. Recommended storage conditions: 2C to 8C.
Related products Page Hybridoma products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .106
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Insect Medium Supplements

4% Agarose Gel
Pluronic F-68, 10% (100X)

Prepared in distilled water. Insect cell culture tested. Recommended storage conditions: 15C to 30C.
Pluronic is a registered trademark of BASF Corporation.

Yeastolate Ultrafiltrate (50X), liquid

Yeastolate Ultrafiltrate is formulated specifically for use with IPL-41 Insect Medium to support the growth of Spodoptera frugiperda (Sf9). Prepared with 200 g/L of yeastolate in distilled water. Yeastolate Ultrafiltrate is ultrafiltered using a 10,000 Da cutoff. Insect cell culture tested. Performance testing: Performance tested in a growth assay using Sf9 cells. Recommended storage conditions: 2C to 8C, in the dark.
Related products Page Insect Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Lactalbumin Hydrolysate, powder (extra soluble)

Tissue-culture grade. Recommended storage conditions: 15C to 30C.
Mycoplasma Detection
Yeast Extraction Solution
Prepared with distilled water. Yeast extract solution is autolyzed at 150 g/L. Recommended storage conditions: 2C to 8C.
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Other Cell Culture Reagents

2-Mercaptoethanol (1,000X), liquid
55 mM in D-PBS. Formula weight: 78. Recommended storage conditions: 2C to 8C.
Trypan Blue Stain

Membrane filtered and prepared in 0.85% saline. Recommended storage conditions: 15C to 30C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Quantity 100 ml Cat. no. 21985-023 Cat. no. 15250-061
Quantity 50 ml
MEM Vitamin Solution (100X), liquid

Prepared in 0.85% saline. The vitamins in this solution are 100X the concentration in a standard MEM Medium. For product formulation please refer to the tables at the end of this chapter. pH: 5.4 to 7.7. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Phytohemagglutinin (M Form) (PHA), liquid

Phytohemagglutinin is a liquid extract of the red kidney bean, Phaseolus vulgaris. This extract is capable of inducing blastogenesis in vitro in various mammalian mononuclear cells. PHA is a crude sodium chloride extract of beans that have been homogenized in saline solution. Performance and quality testing: Each lot is assayed in a performance test system. Recommended storage conditions: 5 to 20C. Attention: This material is hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
D-Glucose (dextrose)
USP grade. Anhydrous C6H12O6 Formula weight: 180. Recommended storage conditions: 15C to 30C.
Sodium Pyruvate Solution 100 mM (100X), liquid

11,004 mg/L. Recommended storage conditions: 2C to 8C. Intended use: In vitro diagnostic (IVD).
Quantity 100 ml 20 100 ml Cat. no. 11360-039 11360-088
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Cell Culture Reagent Formulations

MEM Amino Acids Solution
Base cat. no. Component Amino acids L-Arginine L-Cystine L-HistidineHClH2O L-Isoleucine L-Leucine L-LysineHCl L-Methionine L-Phenylalanine L-Threonine L-Tryptophan L-Tyrosine L-Valine 6,320.00 1200.00 2100.00 2620.00 2620.00 3625.00 755.00 1650.00 2380.00 510.00 1800.00 2340.00 11130 50X liquid, mg/L
MEM Non-Essential Amino Acids Solution

Base cat. no. Component Amino acids L-Alanine L-Asparagine L-Aspartic acid L-Glutamic acid Glycine L-Proline L-Serine 890.00 1320.00 1330.00 1470.00 750.00 1150.00 1050.00 11140 100X liquid, mg/L
G-5 Supplement
Base cat. no. Component Insulin Human transferrin Selenite Biotin Hydrocortisone FGF EGF 17503 g/ml 500.00 5000.00 0.52 1.00 0.36 0.50 1.00
MEM Vitamin Solution

Base cat. no. Component NaCl D-Ca pantothenate Choline chloride Folic acid i-Inositol Nicotinamide Pyridoxal HCl Riboflavin Thiamine HCl 11120 100X liquid, mg/L 8500.00 100.00 100.00 100.00 200.00 100.00 100.00 10.00 100.00
N-2 Supplement
Base cat. no. Component Insulin Human transferrin Progesterone Putrescine Selenite 17502 g/ml 500.00 10000.00 0.63 1611.00 0.52

Base cat. no. Component Arachidonic acid Cholesterol DL--Tocopherol acetate Linoleic acid Linolenic acid Myristic acid Oleic acid Palmitoleic acid Palmitic acid Pluronic F-68 Stearic acid Tween 80 11905 mg/L 2.00 220.00 70.00
10.00 10.00 10.00 10.00 10.00 10.00 100,000.00 10.00 2200.00
Tween is a registered trademark of ICI Americas, Inc. Pluronic is a registered trademark of BASF Corporation.
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new
PrestoBlue Cell Viability Reagent
Save time with an incubation step as short as 10 minutes Obtain high quality results with large assay window and z values Experience convenience with an add-and-read homogenous format Monitor living cells and perform downstream functional assays with no cell lysis
PrestoBlue reagent is a resazurin-based solution that functions as a cell viability indicator by using the reducing power of living cells to quantitatively measure the proliferation of cells. The PrestoBlue reagent contains a cell-permeant compound that is blue in color and virtually nonfluorescent. When added to cells, the PrestoBlue reagent is modified by the reducing environment of the viable cell, turns red in color and becomes highly fluorescent. This change can be detected using fluorescence or absorbance measurements.
Assay Workflow
A 96-well plate containing the cells and the compounds to be tested is prepared. The PrestoBlue reagent is added directly to cells and incubated at 37oC for 10 minutes. The plate is then transferred to a fluorescence reader to measure signal. Results are evaluated by plotting signal versus compound concentration (Figure 1). In addition to this protocol format, the signal can alternatively be detected by absorbance or the assay can be run at room temperature.
PrestoBluetm reagent 1. Add reagent to cells
2. Incubate
3. Read fluorescence
4. Process data
Figure 1. Data obtained using PrestoBlue reagent shows excellent linearity (R2 value of 0.9948) for cells plated at densities between 100 and 100,000 cells/well. The signal for 100 wells was greater than that from zero cells +/- 3 standard deviations, and the Z values were above 0.5 for cell densities of 391 cells/well or greater (Figure 2).
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Reliable, high-quality results in 10 minutes

Figure 2. Excellent data obtained in just 10 minutes. Jurkat cells were plated in cell culture medium in a 384-well plate in quadruplicate starting at 100,000 cells/ well with a two-fold serial dilution. Cells were incubated with PrestoBlue reagent for 10 minutes prior to fluorescence measurements..
Sensitive detection of cell viability

At 10 minutes of incubation, PrestoBlue reagent can detect as few as 98 cells/well, and the signal remains linear over the a broad dilution range. Following a 16-hour incubation with PrestoBlue reagent, as few as 12 cells/well exhibited a signal that was greater than that from zero cells +/3 standard deviations. Z-factor values were above 0.5 for cell densities of 24 cells/well or greater (Figure 3). Note that at low cell densities, the signal-to-background ratio increases after 16 hours incubation.
Product PrestoBlue PrestoBlue Quantity 25ml 100ml Cat. no. A13261 A13262
Figure 3. Extremely sensitive detection. Jurkat cells were plated in cell culture medium in a 384-well plate in quadruplicate starting at 100,000 cells/well with a two-fold serial dilution. Fluorescence measurements were taken following 10 minutes of incubation and following 16 hours of incubation with PrestoBlue reagent.
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Notes:
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Insect Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174 D.Mel-2 Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174 High Five Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174 Sf9 Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175 Sf21 Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175 Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177 293 Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177 FreeStyle 293 Expression Medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 177 CHO-S Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178 DG44 Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178 Primary Human Keratinocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179 Cell Line Development Kits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180 new OptiCHO Express Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180 new OptiCHO Antibody Express Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181 new Gibco Hepatocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182
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Insect Cells
D.Mel-2 Cells
D.Mel-2 Cells (or Schneider S2 insect cells) were isolated from late-stage Drosophila melanogaster (fruitfly) embryos and are commonly used for the transient or stable expression of recombinant proteins. 1 Drosophila cell lines have been historically cultured in stationary systems utilizing T-flasks and serum-supplemented Schneiders Drosophila basal medium.2 Drosophila insect cells are not anchorage dependent and can be readily transferred between monolayer and suspension culture with no significant effect on cell growth; however, we recommend that D.Mel-2 seed stocks be maintained in suspension culture, which allows for more flexibility and ease of use. Prepared from low-passage cells with documented lineage (15 to 20 passages serum-free) 1.5 107 total cells provided Can be thawed and used directly in suspension culture or used as a monolayer for transfection applications Protocols for recovery, culture, and cryopreservation are provided on the product insert Application: For recombinant protein expression. Performance testing: Each lot is tested for growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity confirmed by isozyme analysis and karyotyping. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment procedures as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
High Five Cells
Adapted to Express Five SFM (Cat. no. 10486-025)

High Five Cells (BTI-TN-5B1-4) are a clonal isolate derived from the parental Trichopulsia ni cell line and are commonly used for the expression of recombinant proteins using the Baculovirus Expression Vector System (BEVS). The cells are adapted to serum-free suspension culture in Express Five SFM, which saves significant time and expense associated with the adaptation of cultures to serum-free and/or suspension cultures. Express Five SFM is a protein-free insect cell culture medium optimized to support maximum cell growth and recombinant gene expression. Provided as a frozen stock in Express Five SFM containing 7.5% DMSO 3 106 cells/ml provided Protocols for recovery, culture, and cryopreservation are provided on the product insert Applications: For baculovirus propagation and recombinant protein expression. Performance testing: Each lot is tested for growth and viability post-recovery from cryopreservation and is also tested for the absence of mycoplasmas. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment procedures as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Important Licensing Information: This product is covered by a Limited Use Label License No. 66. By use of this product you accept the terms and conditions of relevant Limited Use Label License. For research purposes only. Limited Use Label License No. 66: High Five Cells: The High Five cell line is patented by the Boyce Thompson Institute for Plant Research, Ithaca, New York, and is covered under U.S. Patent no. 5,300,435. The High Five cell line is sold for research purposes only. Commercial use requires a license from Boyce Thompson. For more information, please contact: Joyce L. Frank, Tel: 607-254-1220; Fax: 607-254-1242; E-mail: jf51@cornell.edu.
References
1. Schneider, I. (1972) J. Embryol. Exp. Morph. 27, 363. 2. Grace, T.D. (1962) Nature 195, 788.
Quantity 1.5 ml Cat. no. 10831-014
Quantity 3 10 cells/ml
6
Cat. no. B855-02
174
Adapted to Sf-900 III SFM (base cat. no. 12658), a protein-free and animal originfree (AOF) medium
Sf9 cells, a clonal isolate derived from the parental Spodoptera frugiperda (fall armyworm) cell line IPLB-Sf21-AE, are commonly used to isolate and propagate recombinant baculovirus stocks and to produce recombinant proteins. Can be scaled up for culture in bioreactorsprovided as a frozen stock in Sf-900 III SFM containing 7.5% DMSO Each lot is tested for growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Storage conditions: 125C to 200C (liquid nitrogen).
Sf9 Cells
Sf9 Cells
Adapted to Sf-900 II SFM (base cat. no. 10902)

Sf9 cells are a clonal isolate derived from Spodoptera frugiperda (fall armyworm) IPLB-Sf21-AE cells and are used for transient or stable expression of recombinant proteins. Sf9 cells are adapted to serum-free suspension culture in Sf-900 II SFM, which saves significant time and expense associated with the adaptation of cultures to serum-free and/ or suspension cultures.1,2 Insect cells have been historically cultured in stationary systems utilizing T-flasks and serum-supplemented basal medium.3 Insect cells are generally not anchorage dependent, and we recommend that the Sf9 seed stocks be maintained in suspension culture, allowing for more flexibility and ease of use. Prepared from low-passage cells with documented lineage (45 to 50 total passages and 15 to 20 passages serum-free) Provided as a frozen stock in Sf-900 II SFM containing 7.5% DMSO 1.5 107 total cells provided Can be thawed and used directly in suspension culture for rapid expansion of cell stocks, propagation of baculovirus stocks, and production of recombinant proteins, or used as a monolayer for transfection or plaque assay applications Protocols for recovery, culture, and cryopreservation are provided on the product insert Applications: For baculovirus propagation and recombinant protein expression. Performance testing: Each lot is tested for growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment procedures as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
Quantity 1.5 107 cells/vial) Cat. no. 12659-017
Adapted to Sf-900 III SFM (base cat. no. 12658), a protein-free and animal originfree (AOF) medium
Sf21 cells, isolated from Spodoptera frugiperda (fall armyworm) ovarian cells, are commonly used to isolate and propagate recombinant baculovirus stocks and to produce recombinant proteins. Can be scaled up for culture in bioreactorsprovided as a frozen stock in Sf-900 III SFM containing 7.5% DMSO Each lot is tested for growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Storage conditions: 125C to 200C (liquid nitrogen).
Sf21 Cells
Quantity 1.5 107 cells/vial) Cat. no. 12682-019
References
1. Smith, G.E., Ju, G., Ericson, B.L., Moshera, J., Lahm, H., Chizzonite, R., and Summers, M.D. (1985) Proc. Natl. Acad. Sci. USA 82, 8404. 2. Vaughn, J.L., Goodwin, R.H., Tompkins, G.L., and McCawley, P. (1977) In Vitro 13, 213. 3. Grace, T.D. (1962) Nature 195, 788.
7 Eukaryotic Cell Lines
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Chapter 7 Eukaryotic Cell Lines Ordering Information

Quantity Cat. no.
Sf21 Cells
Adapted to Sf-900 II SFM (base cat. no. 10902)

Sf21 cells (IPLB-Sf21-AE) were isolated from Spodoptera frugiperda (fall armyworm) ovarian insect cells and are used for transient or stable expression of recombinant proteins, which saves significant time and expense associated with the adaptation of cultures to serum-free and/or suspension cultures.1 Insect cells have been historically cultured in stationary systems utilizing T-flasks and serum supplemented basal medium.2 Insect cells are generally not anchorage dependent and we recommend that the Sf21 seed stocks be maintained in suspension culture, allowing for more flexibility and ease of use. Prepared from low-passage cells with documented lineage (100 to 110 total passages and 15 to 20 passages serum-free) Provided as a frozen stock in Sf-900 II SFM containing 7.5% DMSO 1.5 107 total cells provided Can be thawed and used directly in suspension culture for rapid expansion of cell stocks, propagation of baculovirus stocks, and production of recombinant proteins, or used as a monolayer for transfection or plaque assay applications Protocols for recovery, culture, and cryopreservation are provided on the product insert Applications: For baculovirus propagation and recombinant protein expression. Performance testing: Each lot is tested for growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment procedures as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
1.5 ml
11497-013
References
1. Vaughn, J.L., Goodwin, R.H., Tompkins, G.L., and McCawley, P. (1977) In Vitro 13, 213. 2. Grace, T.D. (1962) Nature 195, 788.
176
Mammalian Cells
293 Cells
Adapted to CD 293 Medium (Cat. no. 11913-019)
293-F and 293-H cells are clonal isolates derived from transformed embryonal human kidney cells. 1,2 The 293-F strain is a fast-growing variant of the 293 cell line. The 293-H strain is a variant of the 293 cell line that demonstrates better adherence in monolayer culture and ease of use for plaque assays and other anchorage-dependent applications. The 293 parental cell line was selected for growth and transfection efficiency.2 Both 293-F and 293-H cell lines can be adapted to serum-free suspension culture in 293 SFM II or CD 293 Medium supplemented with L-glutamine for recombinant protein production and adenovirus propagation. Prepared from low-passage serum-free adapted cells with documented lineage (20 to 25 passages serum-free) Provided as a frozen stock in 293 SFM II containing 7.5% DMSO 7.5 106 total cells provided Can be thawed and used directly in suspension culture for rapid expansion of cell stocks Can be cultured in monolayer for transfection with Lipofectamine 2000 Transfection Reagent 3 Protocols for recovery, culture, and cryopreservation are provided on the product insert Applications: For recombinant protein expression, signal transduction, and receptor binding studies, and adenovirus and adeno-associated virus (AAV) propagation. Performance testing: Each lot is tested for cell growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 2 containment procedures as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.

FreeStyle 293 Expression Medium is a chemically defined, protein-free medium specifically developed for the ability to support the growth and transfection of 293-F cells under suspension-type culture conditions. The medium is a complete, ready-to-use medium that has been supplemented with GlutaMAX-I Supplement and is animal originfree. FreeStyle 293 Expression Medium is able to save significant time and costs associated with adaptation of cell cultures to serum-free conditions. Recommended storage conditions: 2C to 8C, in the dark.
References
1. Graham, F.L., Smiley, J., Russell, W.C., and Nairn, R. (1977) J. Gen. Virol. 36, 59. 2. Harrison, T., Graham, F., and Williams, J. (1977) Virology 77, 319. 3. Ciccarone, V., Chu, Y., Schifferli, K., Pichet, J.P., Hawley-Nelson, P., Evans, K., Roy, L., and Bennett, S. (1999) FOCUS 21, 54 4. Epstein, D.A., Godwin, G.P., Gruber, D.F., and Grefrath, P.I. (1999) FOCUS 21, 22.
Quantity 6 1,000 ml (case) Cat. no. 12338-026
References
1. Graham, F.L., Smiley, J., Russell, W.C., and Nairn, R. (1977) J. Gen Virol. 36, 59. 2. Roy, L., Cates, S., Schifferli, K., Pichet, J.-P., Ciccarone, V., Bennett, S., and HawleyNelson, P. (1999) FOCUS 21, 62. 3. Ciccarone, V., Chu, Y., Schifferli, K., Pichet, J.P., Hawley-Nelson, P., Evans, K., Roy, L., and Bennett, S. (1999) FOCUS 21, 54.
Eukaryotic Cell Lines
Product 293-F Cells 293-H Cells Quantity 1.5 ml 1.5 ml Cat. no. 11625-019 11631-017
Related products Page 293 expression products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
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CHO-S Cells
Adapted to CD CHO Medium (base cat. no. 10743)

CHO-S cells are a clonal isolate derived from Chinese hamster ovary (CHO K1) cells.1 The CHO-S parental cell line was selected for growth and transfection efficiency. 3 The CHO-S cells are adapted to serum-free suspension culture in CD CHO Medium supplemented with L-glutamine and HT Supplement for transient or stable expression of recombinant proteins.2 CHO cells have historically been grown in serum supplemented medium. The use of serum-free media offers ease of downstream purification of expressed product, reduced risk of viral contamination and greater lot-to-lot consistency. CD CHO Medium is a chemically defined, protein-free medium formulated with no components of animal or human origin. Prepared from low-passage cells with documented lineage (20 to 25 passages serum-free) Provided as a frozen stock in CD CHO Medium containing 7.5% DMSO 1.5 107 total cells provided Can be thawed and used directly in suspension culture for rapid expansion of cell stocks Can be transfected in suspension culture using FreeStyle MAX Transfection Reagent Protocols for recovery, culture, and cryopreservation are provided on the product insert See OptiCHO Express and OptiCHO Antibody Express Kits, pages 170 and 171 Applications: For transfection, expression, and large-scale production of recombinant proteins. Performance testing: Each lot is tested for cell growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
DG44 Cells
Preadapted to CD DG44 Medium (Cat. no. 12610-010)*

DG44 cells, dihydrofolate reductase-deficient (DHFR ) a derivative of Chinese hamster ovary (CHO) cells, are routinely used to establish cell lines for production of recombinant protein. They were developed by Dr. Chasin at Columbia University using gamma rays to eliminate the entire DHFR locus. In nonmutant cells, DHFR is an essential enzyme for de novo synthesis of glycine, purines, and thymidylate. This allows DHFR to be used as a dominant selectable marker and a gene amplifier for the expression of proteins in DHFR cell lines. The DHFR mutation in DG44 cells is stable and irreversible, which makes it a suitable mammalian cell line for production of recombinant proteins. GIBCO offers a complete system for the growth and expansion of DG44 (DHFR) cells in suspension, including the DG44 Cells and Media Kit (page 17). Selected for superior cell growth and transfection efficiencies. Applications: For transfection, expression, and large-scale production of recombinant proteins. Performance testing: Each lot is tested for cell growth and viability post-recovery from cryopreservation. The Master Seed Bank has been tested for viruses and mycoplasmas, and sterility and identity have been confirmed by isozyme analysis and karyotyping. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
*CD DG44 Medium is a modification of CD CHO Media.
References
1. DAnna, J.A. (1996) Meth. Cell Sci. 18, 115. 2. Schifferli, K., Jessee, J., and Ciccarone, V. (1999) FOCUS 21, 16.
178
Primary Human Keratinocytes
Adapted to Defined Keratinocyte-SFM (Cat. no. 10744-019)

Primary Human Keratinocytes are derived from normal human tissue and processed and frozen in Defined Keratinocyte-SFM. The cells are effectively adapted to the medium and will establish fully confluent cultures within five days following recovery from cryopreservation when our protocols are followed. The cells are supplied as 1.11.6 106 total cells per vial The lifespan of the cells is performance-tested for 16 population doublings, with most lots demonstrating more than 20 doublings Protocols for recovery and culture are provided on the product insert Application: For experimental protocols requiring monolayer cultures of undifferentiated human keratinocytes. Performance testing: Each lot is tested for cell growth and viability post-recovery from cryopreservation. All lots are screened for Hepatitis B and C, HIV, mycoplasmas, and microbial growth. Recommended storage conditions: 125C to 200C (liquid nitrogen). Caution: Handle material under Biosafety Level 1 containment as defined by the Centers for Disease Control. DMSO has been identified as hazardous. All persons using this product should review the Material Safety Data Sheet before handling.
7 Eukaryotic Cell Lines 179

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Cell Line Development Kits

new
OptiCHO Express Kit
For expression vector construction, high-efficiency transfection of DG44 cells, and development of stable cell lines for protein expression. The OptiCHO Express Kit is designed for efficient growth and transfection of dihydrofolate reductasedeficient (DHFR) Chinese hamster ovary (CHO) DG44 cells in suspension culture. The kit provides a complete workflow protocol as well as the necessary components for efficient transfection of DHFR DG44 cells in a serum-free environment and subsequent stable cell line development, including: pOptiVEC-TOPO vectora bicistronic cloning vector containing a DHFR gene for stable cell line development in CHO-DG44 cells (Figure 1); the vector is TOPO adapted and enables cloning of a PCR product containing your gene of interest with >85% efficiency FreeStyle MAX Transfection Reagentan animal originfree transfection reagent offering high transfection efficiencies and consistent results CD OptiCHO Mediuma chemically defined, protein-free medium (without hypoxanthine or thymidine) specifically designed for highdensity suspension cell culture of recombinant CHODG44 cells and secreted protein yield CD DG44 Mediuma chemically defined, protein-free medium supplemented with hypoxanthine and thymidine to support optimal growth of DG44 cells in suspension culture DG44 cellspreadapted to CD DG44 medium and selected for superior cell growth and transfection efficiencies In addition, the OptiCHO Express Kit provides GIBCO OptiPRO SFM for improved DNA-lipid complex formation, L-glutamine (provided separately for increased media stability), and Pluronic F-68 to control shear forces in suspension cultures. Recommended storage conditions: Pluronic F-68, 15C to 30C; CD DG44 Medium, OptiPRO SFM, CD OptiCHO Medium, and FreeStyle MAX Transfection Reagent, 2C to 8C, in the dark; L-glutamine, pOptiVEC-TOPO vector, CMV Forward Sequencing Primer, and EMCV IRES Reverse Sequencing Primer, 5C to 20C; One Shot TOP10 Chemically Competent E. coli cells, 80C; DG44 cells, liquid nitrogen.
Product Kit components, also available separately pOptiVEC-TOPO TA Cloning Kit CD DG44 Medium FreeStyle MAX Transfection Reagent OptiPRO SFM CD OptiCHO Medium Pluronic F-68, 10% L-glutamine-200 mM (100X)
Quantity 1 kit 1,000 ml 1 ml 100 ml 1,000 ml 100 ml 100 ml
Cat. no. 12744-017 12610-010 16447-100 12309-050 12681-011 24040-032 25030-081
TOPO
CCCTT GGGA
AGGG TTCCC
TOPO
PC
MV
IR ES
4.4 kb
pU
or
TK
pA
Figure 1Map of the pOptiVEC-TOPO cloning vector.
180
DHFR
Am
p i c illi n
C
pOptiVEC-TOPO
new
OptiCHO Antibody Express Kit
Serum-free cell line development and cloning for antibody expression. The OptiCHO Antibody Express Kit is designed for faster serumfree cell line development and cloning for antibody expression in dihydrofolate reductasedeficient (DHFR) Chinese hamster ovary (CHO) DG44 cells in suspension culture (Figure 1). The kit provides a complete workflow protocol and manual, as well as the necessary components for efficient transfection and cloning of DHFR DG44 cells in a serumfree environment and subsequent development of a stable cell line for antibody production. The kit contains: Two expression vectors for light and heavy chainsthe pOptiVECTOPO bicistronic cloning vector and pcDNA3.3 TOPO cloning vector (Figure 2); the vectors are TOPO adapted to enable cloning of a PCR product containing your gene of interest with >85% efficiency FreeStyle MAX Reagentan animal originfree transfection reagent offering high transfection efficiencies and consistent results CD OptiCHO Mediuma chemically defined, protein-free medium (without hypoxanthine or thymidine) specifically designed for high density suspension culture of recombinant CHO DG44 cells and secreted protein yield CD DG44 Mediuma chemically defined, protein-free medium supplemented with hypoxanthine and thymidine to support optimal growth of DG44 cells in suspension culture DG44 cellspreadapted to CD DG44 medium and selected for superior cell growth and transfection efficiencies
In addition, the OptiCHO Antibody Express Kit provides GIBCO OptiPRO SFM for improved DNA-lipid complex formation, Geneticin Selective Antibiotic (G-418) for DHFR selection, L-glutamine (provided separately for increased media stability), and Pluronic F-68 to control shear forces in suspension cultures. Recommended storage conditions: Pluronic F-68, 15C to 30C; CD DG44 Medium, OptiPRO SFM, and CD OptiCHO Medium, 2C to 8C, in the dark; FreeStyle MAX Transfection Reagent, 2C to 8C; Geneticin Selective Antibiotic, 5C to 20C, or 2C to 8C; L-glutamine, pcDNA3.3TOPO TA Cloning Kit, and pOptiVEC-TOPO TA Cloning Kit, 5C to 20C; One Shot TOP10 Chemically Competent E. coli cells, 80C; DG44 cells, liquid nitrogen.
Product Kit components sold separately pcDNA3.3-TOPO TA Cloning Kit pOptiVEC-TOPO TA Cloning Kit CD DG44 Medium FreeStyle MAX Transfection Reagent OptiPRO SFM CD OptiCHO Medium Geneticin Selective Antibiotic, liquid Pluronic F-68, 10% L-Glutamine-200 mM (100X)
Quantity 1 kit 1 kit 1,000 ml 1 ml 100 ml 1,000 ml 100 ml 100 ml 100 ml
Cat. no. K8300-01 12744-017 12610-010 16447-100 12309-050 12681-011 10131-027 24040-032 25030-081
Current steps
Adaptation 3 months Expression vectors 3 months CHO cells 12 months Stable polyclonal populations 12 months Ampli cation 1 round 23 months Clone selection 3 months Adaptation 3 months Protein production cell lines
OptiCHO cell engineering

Adaptation 0 months Expression vectors 0.5 months CHO cells 1 month Stable polyclonal populations 1 month Ampli cation 1 round 23 months Clone selection 0 months Adaptation 3 months Protein production cell lines
7 Eukaryotic Cell Lines
Total time: 1619 months
Total time: 79 months
Potential for more than 6 months time saved using OptiCHO Express Kits
Figure 1Time savings throughout the cell line development process.
Figure 2Maps of the pOptiVEC-TOPO and pcDNA3.3-TOPO cloning vectors.
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new
Gibco Hepatocytes
Gibco Human Cryopreserved Hepatocytes, Transporter Qualified
Contain functional membrane receptors and transporters Facilitate effective transporter uptake and basolateral efflux (plated) experiments Have stringent release specs: 80% viability and 80% confluency (plated cells)
Primary hepatocytes isolated from the liver are effective tools for the in vitro evaluation of metabolism, drug-drug interactions, hepatotoxicity, and transporter activity. Gibco hepatocytes have high viabilities and in vivo-like enzyme expression levels. Extensive selection of lots Viabilities routinely >80% Characterized for phase I and phase II drug metabolizing enzyme activities Multiple large lots available - ideal for reproducible long-term and multi-site studies
Gibco Human Hepatocytes, Induction Qualified

Prequalified for CYP1A2, CYP2B6 and CYP3A induction 80% viability and 80% confluency (if plated cells) Minimum specific activities: 10-fold induction of CYP1A2 5-fold induction of CYP2B6 3-fold induction of CYP3A4
Gibco Cryopreserved Human and Animal Hepatocytes

At Invitrogen, our hepatocyte isolationists are extensively trained in proper techniques to ensure optimal cell health. As a result, gibco hepatocytes have high viabilities, in vivo-like enzyme expression levels, and if released as plateable cells, excellent confluencies that contribute to polarization and functioning cell-cell contacts Extensive selection of lots Viabilities routinely >80% Characterized for phase I and phase II drug metabolizing, enzyme activities Multiple large lots availableideal for long-term studies across sites Characterization and quality control: 10-point cell morphology check backed by photomicrographscell membrane integrity, organelle size, presence of lipid droplets, nucleus size and shape, cytosolic clarity, cell shape, level of cell debris, cell excretion products, cell-cell contacts (plateable cells only)and reestablishment of bile canalicular networks (plateable cells only) Metabolic activity testingECOD, 7-HCG, 7-HCS; human hepatocytes includeCYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 CYP3A, FMO Application qualification testsattachment efficiency, monolayer confluency fold induction with prototypical inducers, transporter uptake and efflux, in situ intrinsic clearance Additional characterization datagenotyping, optimal seeding densities viability stabilities, donor demographics
Gibco Human Cryopreserved Hepatocytes, Plated Metabolism Qualified

Useful for the assessment of intrinsic clearance (CLint) in low turnover compounds Prequalified according to CLint of midazolam, tolbutamide, dextromethorphan 80% viability and 75% attachment efficiency
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Product Human cryopreserved plateable hepatocytes (Each lots certificate of analysis has an exact cell count.) Human Cryopreserved Plateable Hepatocytes, Transporter Qualified Human Cryopreserved Plateable Hepatocytes, Transporter Qualified Human Cryopreserved Plateable Hepatocytes, Induction Qualified Human Cryopreserved Plateable Hepatocytes, Induction Qualified Human Cryopreserved Plateable Hepatocytes, Metabolism Qualified Human Cryopreserved Plateable Hepatocytes, Metabolism Qualified Human cryopreserved suspension hepatocytes Human Cryopreserved Suspension Hepatocytes, Transporter Qualified Human Cryopreserved Suspension Hepatocytes, Transporter Qualified Human Cryopreserved Suspension Hepatocytes, Metabolism Qualified, Male Human Cryopreserved Suspension Hepatocytes, Metabolism Qualified, Male Human Cryopreserved Suspension Hepatocytes, Metabolism Qualified, Female Human Cryopreserved Suspension Hepatocytes, Metabolism Qualified, Female Human Cryopreserved Suspension Hepatocytes, Metabolism Qualified, Pooled Donors Human Cryopreserved Suspension Hepatocytes, Metabolism Qualified, Pooled Donors 48 million viable cells 912 million viable cells 48 million viable cells 912 million viable cells 48 million viable cells 912 million viable cells 48 million viable cells 912 million viable cells HMCSTS HMCSTL HMCS1S HMCS1L HMCS2S HMCS2L HMCS3S HMCS3L 48 million viable cells 912 million viable cells 48 million viable cells 912 million viable cells 48 million viable cells 912 million viable cells HMCPTS HMCPTL HMCPIS HMCPIL HMCPMS HMCPML Quantity Cat. No.
Animal suspension and plateable cryopreserved hepatocytes (Please inquire for minipig, guinea, and other custom species.) Dog (Beagle) Cryopreserved Hepatocytes, Male Dog (Beagle) Cryopreserved Hepatocytes, Female Dog (Beagle) Cryopreserved Hepatocytes, Plateable Male Dog (Beagle) Cryopreserved Hepatocytes, Plateable Female Fish (Rainbow Trout) Cryopreserved Hepatocytes, Male Fish (Rainbow Trout) Cryopreserved Hepatocytes, Female Fish (Rainbow Trout) Cryopreserved Hepatocytes, Mixed Gender Monkey (Cynomolgus) Cryopreserved Hepatocytes, Male Monkey (Cynomolgus) Cryopreserved Hepatocytes, Female Monkey (Cynomolgus) Cryopreserved Hepatocytes, Plateable Male Monkey (Cynomolgus) Cryopreserved Hepatocytes, Plateable Female Mouse (CD-1) Cryopreserved Hepatocytes, Male Mouse (CD-1) Cryopreserved Hepatocytes, Female Mouse (CD-1) Cryopreserved Hepatocytes, Plateable Male Mouse (CD-1) Cryopreserved Hepatocytes, Plateable Female Rabbit (New Zealand White) Cryopreserved Hepatocytes, Male Rabbit (New Zealand White) Cryopreserved Hepatocytes, Female Rabbit (New Zealand White) Cryopreserved Hepatocytes, Plateable Male Rabbit (New Zealand White) Cryopreserved Hepatocytes, Plateable Female Rat (Sprague-Dawley) Cryopreserved Hepatocytes, Male 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells 48 million viable cells DGCS10 DGCS20 DGCP10 DGCP20 TRCS10 TRCS20 TRCS30 MKCS10 MKCS20 MKCP10 MKCP20
MSCS10 MSCS20
Eukaryotic Cell Lines
MSCP10 MSCP20 RBCS10 RBCS20 RBCP10 RBCP20 RTCS10
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Gibco Fresh Hepatocytes

Reliable supply from the industrys largest sourcing network Stringent release specs: 80% viability and 80% confluency (plated cells) Ready for useplated to your specifications, or provided in suspension Primary human hepatocytes, under appropriate culture conditions, model metabolism, induction, and inhibition similar to the in vivo liver. Fresh human hepatocytes are used to test potential therapeutic compounds for their metabolic stability, metabolic profile, probability of hepatotoxicity, and ability to inhibit or induce metabolic enzymes and hepatic transporters. To achieve optimal in vitro results, primary hepatocytes must be carefully isolated to preserve their drug-metabolizing enzymes, transporters, and cellular activities. Our hepatocyte isolationists are experts in proper techniques to ensure optimal cell health. As a result, our fresh hepatocytes have high viabilities, excellent confluencies and in vivo-like enzyme expression levels.
Rapid Alert notifications

Rapid Alert emails notify you right away about the impending availability of fresh primary human hepatocytes. When you receive a Rapid Alert email, just decide if you need hepatocytes, and contact us to place an order. Each Rapid Alert email contains: a summary of the donors demographics (gender, age, race, BMI, and history of alcohol, drugs, or medications) plate and suspension formats available for order a shipping date for the hepatocytes a phone number and email to place your order or request additional information. To sign up for Rapid Alert, access our online form at www.invitrogen.com/hepatocytes
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Chapter 8 Animal Sera

Fetal Bovine Sera (FBS) and other Sera for Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186 General Animal Sera Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186 The iMATCH MPA Sera Lot Matching Tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186 FBS Selection Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187 Fetal Bovine Sera (FBS). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187 qualified . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188 qualified, heat-inactivated . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188 qualified, gamma irradiated . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189 Specialty FBS Selection Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189 dialyzed. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189 new charcoal stripped. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189 ES cell qualified . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189 new MSC qualified . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190 ultra-low IgG FBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190 Other Sera Selection Guide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190 Newborn Calf Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Newborn Calf Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Newborn Calf Serum, heat-inactivated . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Bovine Sera. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Bovine Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Bovine Serum, heat-inactivated . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Donor Bovine Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Donor Bovine Serum, with iron . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 Horse Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Horse Serum. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Horse Serum, heat-inactivated . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Other Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Chicken Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Goat Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Lamb Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Porcine Serum. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 Rabbit Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192 FBS Quality Control Test Comparison. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193 Other Sera Quality Control Test Comparison . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194 Final Product Quality Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 196 Performance Tests: Fetal Bovine Sera (FBS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197 Special Proccessing of Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 200
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Fetal Bovine Sera (FBS) and other Sera for Cell Culture
A History of Value and Trust: A World leader Today
Global sourcing capabilities, multiple aseptic-processing facilities worldwide, batch size capabilities up to 2,000 litres, State-of-the-Art processing (including steam sterilisation in place/true pool technology), an unsurpassed commitment to both product quality and traceability, and a deep understanding of customers requirements, are just a few of the factors which have contributed over four decades to establishing Life Technologiestm as the major supplier of Fetal Bovine Serum (FBS) and other animal sera in the world today.
The iMATCH MPA Sera Lot Matching tool

iMATCH is a tool that our FBS specialists use to find you the most consistent or highest performing lot of serum available. iMATCH uses a proprietary algorithim conducting multiparametric analysis of Certificate of analysis performance parameters to match your needs. The iMATCH ser vice is free of charge, only available through Life Technologiestm and; Saves time by reducing or eliminating serum testing Makes serum choices based on performance, not source Minimizes fluctuations in cell culture performance Increases confidence in you search results iMATCH is our promise to give you the best serum batch for your application every time.
Vertical Integration of gibco Sera Production

Life Technologiestm oversees the preparation of gibco sera through its entire process. Using specialised equipment, Standard Operating Procedures (which cover every step from blood collection to final product validation) and ongoing internal audits. Life Technologiestm ensures that quality is engineered into the product at every stage. This control over the entire process results in a consistent, high quality product with lower lot-to-lot variation.
Shipping and Storage

(a) All serum is routinely packed in dry ice for shipment. (b) We recommend that you store serum at -5C to -20C and use it before the expiration date indicated on the label. The plastic bottles may be stored at temperatures as low as -60C to -70C. (c) We also recommend that you avoid multiple freeze-thaws. If the serum is likely to be frozen and thawed frequently, dispense it aseptically into sterile containers in appropriate volumes and store it frozen in these containers. It is recommended that serum is thawed at +4C to minimaise precipitation.
Geographical Differentiation of FBS

In response to the demand from the research community and biopharmaceutical companies. Life Technologiestm offers a choice of FBS differentiated by source. Our FBS product line feature products obtained from countries meeting USDA import requirements and/or E.U. import rules. All FBS products meet our rigorous quality control criteria. This differentiation allows the user to make an informed purchasing choice and offers price flexibility
Handling of Bovine Origin Materials

As with all materials of biological origin, Life Technologiestm recommends that all bovine products are treated as potentially harmful and that appropriate care is taken when handling and disposing of them.
Manufacture in Country of Origin

In response to requests from the biopharmaceutical industry, Life Technologiestm now final-processes all normal sera of Australian, New Zealand and USA origins in the actual country of origin.
Intended Use of Products

The products listed in this chapter are for research use only, except where otherwise denoted. Research use only products are not intended for diagnostic use. The safety and efficacy of these products in diagnostic or other clinical uses has not been established. Products listed for in vitro diagnostic use are indicated by the symbol IVD and are medical devices subject to the requirements of Directive 98/79/EC of the European Parliament and of the Council of 27 October 1998 on in vitro diagnostic medical devices [Official Journal L 331 of 07.12.1998] and to the requirements of the Code of Federal Regulations Title 21, Part 820: Quality Systems Regulation. These products may also be used for research applications, and in appropriate circumstances, as raw material components in further manufacturing applications. It is the end users responsibility to qualify these products for their specific application. They have not been qualified nor are they intended for any animal or human therapeutic uses.
Label information will change to incorporate the CE mark on for in vitro diagnostic use products, and a glossary of other symbols found on the labels can be found on page 237.
Traceability: your Proof of Serum Quality

The Certificate of Analysis available for each lot of gibco serum identifies the country of origin. This information is fully supported by extensive documentation that certifies each lot from collection through processing, testing, and delivery. This traceability is your proof of serum quality.
Batch Testing with Reserves

Many of our customers purchase serum without pretesting with satisfactory results. However, we recognise the fact that some cell types are particularly sensitive to batch to batch variations in serum. Reserve testing allows customers to obtain a sample for testing in their own application with a reserve held on that batch. Serum is normally held on reserve for four weeks. If a longer test period is required, please advise us at time of reserve request.
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FBS Selection Guide

Product Certified FBS Description/usage guidelines Undergoes special biochemical/hormonal profile and bacterial tests. Guaranteed low endotoxin (10 EU/ml) and low hemoglobin (15 mg/dl) levels Use with your most sensitive cells Heated for 30 min at 56C with mixing to inactivate complement Especially suited for immunological work Our most popular FBS product. High quality and exceptional value Suitable for general applications, especially those that do not require defined biochemical/hormonal profiles or bacteriophage testing Endotoxin 50 EU/mllevels routinely less than 10 EU/ml Hemoglobin 25 mg/dl Exceptions for FBS EU Approval origins. Endotoxin <100 EU/ml and Hemoglobin<30 MG/dl (South America only) Qualified, heat-inactivated FBS Heated for 30 min at 56C with mixing to inactivate complement Especially suited for immunological work Countries that meet EU importation requirements (excluding South America) United States Australia Countries that meet EU importation requirements (South America only) Countries that meet EU importation requirements (excluding South America) Qualified Gamma Irradiated FBS Intended for application where virus contamination is of concern. Gamma Irradiated with a minimum dose of 25kGys. Intended for applications where virus contamination is of concern Countries that meet EU importation requirements (South America only) Countries that meet EU importation requirements (excluding South America) 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 10106-151 10106-169 16140-063 16140-071 10100-139 10100-147 10500-056 10500-064 10108-157 10108-165 10499-036 10499-044 10109-155 10109-165 Source United States Quantity 100 ml 500 ml Cat. no. 16000-036 16000-044
Certified, heat-inactivated FBS Qualified FBS
United States United States Australia Countries that meet EU importation requirements (South America only)
100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml
10082-139 10082-147 26140-087 26140-079 10099-133 10099-141 10270-098 10270-106
Fetal Bovine Sera (FBS)

Fetal Bovine Serum, certified
Endotoxin level: 10 EU/ml. Hemoglobin level: 15 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Fetal Bovine Serum, certified, heat-inactivated

Endotoxin level: 10 EU/ml. Hemoglobin level: 15 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Animal Sera 187
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Fetal Bovine Serum, qualified

Endotoxin level: 50 EU/ml (levels routinely 10 EU/ml). Hemoglobin level: 25 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Fetal Bovine Serum, qualified, heat-inactivated


Origin: Australia

Origin: Australia

Origin: Countries meeting EU importation requirements
Endotoxin level: 100 EU/ml (levels routinely less than 20 EU/ml). Hemoglobin level: 30 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).

Origin: Countries meeting EU importation requirements
Endotoxin level: 100 EU/ml (levels routinely 20 EU/ml). Hemoglobin: 30 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Quantity 100 ml 500 ml 100 ml 500 ml Cat. no. 10270-098 10270-106 10106-151 10106-169
Quantity 100 ml 500 ml 100 ml 500 ml Cat. no. 10500-056 10500-064 10108-157 10108-165
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Fetal Bovine Serum, qualified, gamma irradiated

Origin: Countries meeting EU importation requirements stem cells.
Specialty FBS Selection Guide

Product Dialyzed FBS Description/usage guidelines Dialyzed by tangential flow filtration utilizing 10,000 MW cutoff filters Performance tested for cloning and plating efficiency Ideal for radiolabeling assays IgG levels are less than 5 g/ml, and the BVD antibody titer is low or not detectable Suitable for antibody production and veterinary applications Specially tested for the ability to sustain undifferentiated cellular morphology of embryonic stem cells Crucial for the successful maintenance of embryonic stem cells Performance tested using a standard 14-day MSC CFU-f assay Each lot is tested against an in-house FBS reference standard using cells from a master cell bank of MSCs from normal bone marrow donors. This ensures lot-to-lot performance consistency. A serum product that will reduce lot-to-lot variability in hormone levels, which will help eliminate some of the influences steroids and other components have on cells Ideal for customers doing cell culture work using immunoassay systems and insulin assay methods Source United States Quantity 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml Cat. no. 26400-036 26400-044 16250-086 16250-078 16141-061 16141-079 12662-011 12662-029
Ultralow-IgG FBS
United States
ES cell-qualified FBS
United States
new MSC Qualified FBS
Countries that meet USDA importation requirements (Mexico, Central America)
new Charcoal Stripped

FBS
Countries that meet USDA importation requirements (Mexico, Central America)
50 ml 500 ml
12676-011 12676-029
Fetal Bovine Serum, dialyzed

new
Fetal Bovine Serum, charcoal stripped

Endotoxin level: 50 EU/ml (levels routinely 10 EU/ml). Hemoglobin: 25 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: For research use only.
Fetal Bovine Serum, ES cell qualified

Origin: United States. Specially tested for the ability to sustain undifferentiated cellular morphology of embryonic stem cells.
Hemoglobin level: 25 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Animal Sera 189
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Fetal Bovine Serum, MSC qualified
Fetal Bovine Serum, ultra-low lgG FBS

lgG levels are less than 5 g/ml, and the BVD antibody titer is low or not detectable. Endotoxin level: 50 EU/ml (levels routinely less than 10 EU/ml). Hemoglobin level: 25 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).

Specifically tested for the ability to attain enhanced mesenchymal stem cell clonal efficiency. Endotoxin level: 50 EU/ml (levels routinely 10 EU/ml). Hemoglobin level: 25 mg/dl. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Related products Page Stem cell research products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .117
Other Sera Selection Guide

Product Newborn Calf Serum Newborn Calf Serum, heat-inactivated Bovine Serum Bovine Serum, heat-inactivated Donor Bovine Serum Donor Bovine Serum, with iron Horse Serum Horse Serum, heat-inactivated Chicken Serum Goat Serum Lamb Serum Porcine Serum Rabbit Serum Description/usage guidelines Collected from calves typically 20 days old or younger Collected from calves typically 20 days old or younger Heated for 30 min at 56C with mixing to inactivate complement Collected from prime cattle with an age range of 1236 months, but usually 24 months or younger Collected from prime cattle with an age range of 1236 months, but usually 24 months or younger. Heated for 30 min at 56C with mixing to inactivate complement Collected from donor cattle with an age range of 1236 months, but usually 24 months or younger Collected from donor cattle with an age range of 1236 months, but usually 24 months or younger. Ferric citrate complex added at 6.0 mg/dl EIA tested (negative) from a donor herd EIA tested (negative) from a donor herd Heated for 30 min at 56C with mixing to inactivate complement Collected from healthy birds slaughtered for human consumption Collected from donor herd Collected from animals younger than 1 year old Collected from animals younger than 1 year old Collected from animals typically 6 months old or younger Source New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand New Zealand United States Quantity 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 100 ml 500 ml 500 ml 500 ml 100 ml 500 ml 100 ml 500 ml 500 ml 100 ml 500 ml 500 ml 500 ml 100 ml 500 ml Cat. no. 16010-167 16010-159 26010-066 26010-074 16170-086 16170-078 26170-035 26170-043 16030-074 10371-029 16050-130 16050-122 26050-070 26050-088 16110-082 16210-064 16210-072 16070-096 26250-084 16120-099 16120-107
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Newborn Calf Sera

Newborn Calf Serum
Origin: New Zealand
Collected from calves 20 days old or younger. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Newborn Calf Serum, heat-inactivated

Origin: New Zealand
Collected from calves 20 days old or younger. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Bovine Sera (formerly Calf Sera)

Bovine Serum
Origin: New Zealand
Collected from cattle 1236 months of age. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Bovine Serum, heat-inactivated

Origin: New Zealand
Donor Bovine Serum

Origin: New Zealand
Donor Bovine Serum, with iron

Origin: New Zealand
Ferric citrate complex at 6.0 mg/dl. Collected from cattle 1236 months of age. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Cat. no. 16030-074
Quantity 500 ml
8 Animal Sera 191
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Horse Sera
All GIBCO horse sera have been tested and found negative for equine infectious anemia (EIA).
Horse Serum
Origin: New Zealand
EIA tested (negative) serum from a donor herd. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Horse Serum, heat-inactivated

Origin: New Zealand
EIA tested (negative) serum from a donor herd. Recommended storage conditions: 5C to 20C. Intended use: In vitro diagnostic (IVD).
Other Sera
Chicken Serum
Origin: New Zealand
Porcine Serum
Origin: New Zealand
Goat Serum
Origin: New Zealand
Rabbit Serum
Origin: United States, New Zealand
Lamb Serum
Origin: New Zealand
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FBS Quality Control Test Comparison

Certified 16000 Certified heatinactivated 10082 Qualified 26140,10099, Qualified heat-inactivated 16140, 10100
Test Bacteriophage tested Biochemical profile Bovine IgG (<5 g/ml) Determination of % oxy Hb (>70%) Electrophoretic pattern Endotoxin (EU/ml) ES cell performance assays MSC CFU-f assay Glucose (<5 g/ml) Hemoglobin (mg/dl) Hormone profile Mycoplasmas Osmolality Performance tested pH Sf9 cell assay Sterility testing (bacteria/fungi) Tetracycline* Total protein Viral testing Akabane virus Bluetongue virus Bovine adenovirus FA Bovine parvovirus FA Bovine respiratory syncytial virus FA Bovine viral diarrhea virus FA Bovine viral diarrhea neutralization assay Cytopathic agents (e.g., IBR) Hemadsorbing agents (e.g., PI-3) Reovirus FA Rabies virus FA
Qualified 10270, 10106
Qualified heat-inactivated 10500, 10108
Dialyzed 26400
Ultralow-IgG 16250
Charcoal Stripped 12676
ES Cell-Qualified 16141
MSC Qualified 12662
Animal Sera 193
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Other Sera Quality Control Test Comparison

Base cat. no. 16010 Newborn Calf Serum Cytotoxicity assay Electrophoretic pattern Endotoxin (EU/ml) Hemoglobin (mg/dl) Mycoplasmas Osmolality pH Sp2 performance assay Sterility testing (bacteria/fungi) Total protein VERO performance assay Viral testing Bluetongue virus Bovine adenovirus FA Bovine parvovirus FA Bovine respiratory syncytial virus FA Bovine viral diarrhea virus FA Cytopathic agents EIA coggins Equine herpes virus Equine viral arteritis Hemadsorbing agents Porcine adenovirus Porcine hemagglutinating encephalitis Porcine parvovirus Rabies FA Reovirus FA Transmissible gastroenteritis virus 26010 Newborn Calf Serum, heat-inactivated 16170 Bovine Serum 26170 Bovine Serum heat-inactivated 16030 Donor Bovine Serum 10371 Donor Bovine Serum with iron
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16050 Horse Serum
26050 Horse Serum, heat-inactivated
16110 Chicken Serum
16210 Goat Serum
16070 Lamb Serum
26250 Porcine Serum
16120 Rabbit Serum
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Final Product Quality Control

A representative number of quality control samples from each production lot are analyzed using the following methods: Chemical checks. Osmolality is measured using freezing point depression to verify compliance with product specifications. All osmometers are calibrated daily, using standard solutions. pH is measured to verify compliance with product specifications. All pH meters are calibrated daily using standard solutions. Electrophoretic profiles are monitored to confirm serum identity and integrity. Samples are applied to a cellulose acetate medium and allowed to migrate in a buffer system. The strips are stained, cleared, dried, and scanned using a densitometer to determine relative concentrations of albumin and globulin fractions. To verify that proper collection and processing procedures have been used, hemoglobin is measured spectrophotometrically using a modified Fleming method 1 for oxyhemoglobin determination. As the age of the animal increases, the total protein increases proportionately. To confirm animal age and verify compliance with product specifications, total serum protein is measured using an automated Biuret method. The absorbance of a sample combined with the Biuret reagent is measured at various wavelengths using a colorimeter. Results are automatically calculated and compared to a standard curve, and protein values are calculated in grams per decilitre. Stability testing program. The expiration date shown on every in vitro diagnostic (IVD)-labeled product is an indication of how long the product will deliver effective performance. Our stability program establishes and verifies product expiration dates. Product groups are monitored at periodic intervals to determine the length of time that the product will deliver acceptable performance under recommended storage conditions. Microbiological tests. The absence of bacterial and fungal contamination is confirmed using methods referenced in the current edition of the U.S. Pharmacopeia (USP) and/or the European Pharmacopoeia. All sera are tested for the absence of mycoplasmas using the largevolume method of Barile and Kern. 2 The test is accurate within the detection limits of the methods referenced. The samples are pooled and cultured in broth for a minimum of three weeks, with no fewer than four subcultures onto mycoplasma agar plates. The plates are incubated at 36C 2C under both aerobic and anaerobic (95% nitrogen, 5% CO2) conditions. The agar plates are examined microscopically for growth at weekly intervals during the test procedure. Dienes stain is used to detect mycoplasma colonies in agar plates that are suspected of contamination. The plates then are reexamined microscopically. Inoculated broth bottles are periodically monitored for turbidity and evidence of pH shift. All bovine sera of Australian , New Zealand, and USA Origins are screened for noncultivatable mycoplasmas using the Hoechst fluorescent DNA stain 3 to detect the presence of mycoplasmas that are noncultivatable by the broth method, including species of M. hyorhinis, within the detection limits of the test method referenced. Viral tests. Adventitious agents are detected using procedures from the Code of Federal Regulations, Title 9 CFR, Part 113.53, Requirements for Ingredients of Animal Origin.
Bovine cell cultures previously demonstrated to be free of known adventitious agents are passaged for three subcultures in growth medium containing 15% test serum for 21 days. Negative reference cultures are maintained in parallel, using control medium and serum previously demonstrated as negative for adventitious agents. All cultures are examined throughout this period for evidence of virus-induced morphological changes or cytopathogenic effects. At day 14 of a 21-day culture period, parallel culture flasks containing test serum are evaluated by standard virus-testing protocols, as follows. A flask of test cells is trypsinized, and cells are added to six chamber slides, with a total surface area of 6 cm2. Negative reference slides are prepared in parallel. Individual positive controls are prepared by adding strains of bovine viral diarrhea virus (BVDV), and for certain sera as detailed in the Quality Control Text Comparison, bovine parvovirus, bovine adenovirus, rabies virus, and reovirus to individual chamber slides of test cultures. After an additional seven days of culture (21 days total), viruses are detected by fluorescent antibody techniques. Cytopathogenic effects (CPE) or other virus-induced effects are observed through hematoxylin and eosin staining of test cultures. Cells are examined for presence of CPE, inclusion bodies, syncytia formation, or other abnormal effects. Cells from a second flask are tested for hemadsorption (HAD) of human type O, guinea pig, and day-old chick erythrocytes, following a procedure patterned after 9 CFR 113.46(b). The test is conducted at 2C to 8C and 20C to 24C. Negative control cells previously infected with PI-3 virus serve as positive controls. Uninfected cells are used as the negative control. These tests detect the presence of viral contamination within the detection limits of the methods referenced. With the exception of noncytopathic BVDV in Fetal Bovine Serum,cell batches of serum must test negative for virus before release for sale. Endotoxin test. The Limulus Amoebocyte Lysate (LAL) gel-clotting procedure is used to measure endotoxin values in all FBS 1 with the exception of FBS and other blood and sera products manufactured in New Zealand. New Zealand uses a chromogenic kinetic test procedure which measures the concentration of endotoxin spectrophotometrically, based on the initial rate of reaction.4
References
1. Fleming, A.F. and Woolf, A.J. (1965) Clin. Chem. 12, 67. 2. Barile, M.F. and Kern, J. (1971) Proc. Soc. Exp. Biol. Med. 138, 432. 3. Chen, J.R. (1977) Exp. Cell Res. 104, 255. 4. Current USP.
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Performance Tests: Fetal Bovine Sera (FBS)

The most important criterion in selecting FBS is its ability to support the growth of particular cell types. Therefore, in addition to ensuring that each lot of FBS passes our stringent sterility control tests, we simulate laboratory conditions to test each lot for three important performance character istics: cloning efficiency, plating efficiency, and diploid fibro blast growth promotion. 1 Our exclusive performance testing procedures provide quality sera that are defined in terms of their suitability for these three applications. These tests help ensure that each lot of FBS we select for you will provide optimal results in your application. Cloning efficiency assay. Cat No`s 10106, 10108, 10109, 10270, 10500 The cloning efficiency assay analyses the ability of each batch to support cloning and growth of myeloma cells and derived hybridomas. Cells used to determine cloning efficiency are Sp2/O-Ag14 or P3x63 Ag8.653. These are routinely maintained at log phase RPMI 1640 Medium supplemented with 10% Foetal Bovine Serum. The Serum is assayed in microtitre plates and the demanding one cell per well test simulates laboratory conditions for selection of a single hybrid. Cloing assay results are normalised relative to an established reference serum. The cell suspension being used is adjusted to the desired concentration of 5 cells/ml by dilution with RPMI 1640 Medium supplemented with 20% test or reference serum. The cell suspensions are then swirled gently to evenly distribute the cells before dispensing 200 l into each well of a 96 well microtitre plate thus giving a final concentration of 1 cell per well. Cloning efficiency is calculated as follows: cloning efficiency = average number of positive wells total number of wells inoculated 100
Plating Efficiency Assay Cat No`s. 10106, 10108, 10109, 10270, 10500 The plating efficiency assay determines the ability of a particular batch of FBS to grow a continuous adherent cell line from a very low inoculum density. BHK-21 cells are selected for use in the assay because of their ability to detect subtle variations in serum ability to support cellular attachment and proliferation. BHK-21 cells are maintained in logarithmic phase at + 37 oC in Glasgow MEM supplemented with 10% Newborn Calf serum and 10% Trytose Phosphate Broth. Nutrient Mix F12 (HAM) supplemented with 1% glutamine is used as growth medium. Five flasks of growth medium supplemented with 20% test serum are inoculated at 40 cells/ml. After incubation for 7 days at + 37OC and 5% CO2, in a humidified incubator, the cells in the flasks are stained with Giemsa and the cell colonies counted. Results are normalised relative to a previously qualified reference serum as follows: Plating Efficiency PE = no of Colonies Counted No of Cells Inoculated 100
Relative Plating Efficiency (RPE) is calculated as: RPE = PE in Test Serum PE in control serum
The relative cloning efficiency (RCE) as follows: RCE = cloning efficiency of test serum cloning efficiency of reference serum Cloning Efficiency Assay. Cat No`s. 16000, 10082, 26140, 16140, 10099, 10100 The cloning efficiency assay analyses the ability of each FBS lot to support cloning and growth of murine The following cell lines are validated for use in this assay: Sp2/O-Ag14 ( ATCC #CRL 1581) P3x63-Ag8.653 (ATCC #CRL 1580) Each FBS lot is assayed in duplicate microtitre plates at two serum concentrations and at two cell innocula (10% serum and one cell/well 4% serum and 5 cells/well). Cloning assay results are normailsed relative to an established reference serum. The higher serum concentration is representative of many routine procedures in hybrid selection, while the lower serum concentration is a more stringent test designed to augment detection of subtle variations among serum lots. The remaining 1-cell/ well test simulates laboratory conditions for selection of a single hybrid. The certificate of analysis, provided with each lot of FBS, reports cloningefficiency as an average of the two test conditions.
Plating Efficiency Assay Cat No`s. 16000, 10082, 26140, 16140, 10099, 10100 The plating efficiency assay uses a human transformed cell line to determine the suitability of each lot for growing continuous adherent cell lines. The cell line validated for determining plating efficiency, A549 (Human Lung Carcinoma, ATCC #CCL, 185), was selected because of it`s ability to support cellular attachment and proliferation. The A549 cell line is maintained at 36oC 2o C at subconfluent density in Dulbecco`s Eagle Medium (D-MEM) supplemented with 10% FBS. Each FBS lot is tested in triplicate at two serum concentrations and at two cellular inocula (10% serum and 100 cells/well, 4% serum and 200 cells/well. After incubation for approximately 10 to 14 days at + 36oC 2o C in a humidified CO2 incubator, the plating efficiency is determined by counting cell colonies. Results are normalized relative to the reference control serum. The two serum concentrations demonstrate the lot`s ability to support growth at both reduced and standard levels. The certificate of analysis, provided with each lot of FBS, reports plating efficiency values as an average of the last two test conditions.
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Diploid fibroblast growth promotion assay. The growth promotion assay measures the ability of each FBS lot to support proliferation of fastidious human diploid fibroblasts through multiple subcultures. The WI-38 cell line (human diploid lung fibroblast, ATCC # CCL 75) is validated for use in this assay. WI-38 cells are inoculated into duplicate 25 cm2 flasks at low density (2 105/flask) with 5% FBS and propagated through three sequential 7-day subcultures. At each passage, the cells are subcultured to the original cell inoculation density. The stressful split ratio, number of population doublings, and reduced serum concentration are stringent measurements of the serum lots ability to grow fastidious diploid cells. The three sequential subcultures ensure a true measurement of the test lots growthpromoting ability by minimizing any residual effects of the previous growth medium. The Certificate of Analysis provided with each lot of FBS reports the average number of cells per duplicate flask from the third subculture. Test values are normalized relative to the reference control. Diploid fibroblast growth assay methodology. The growth promotion assay is performed as follows: 1. Growth medium supplemented with 5% test serum or a previously qualified reference serum is prepared. The basal growth medium is HBME:EBME (1:1) containing L-glutamine. Fresh 5% serumsupplemented growth medium is prepared for each fluid change and subculture during the assay, as required. 2. WI-38 cells (2 105) at a low passage level are inoculated into each flask containing 9.5 ml of the appropriate growth medium in duplicate 25 cm2 flasks. The flasks are maintained loosely capped in a 46% CO2, 36C 2C atmosphere for the initial 24 hours. The flasks then are capped tightly and incubated for 7 days at 36C 2C, with complete medium replenishment at day 2 to day 3. 3. Cells in each of the duplicate flasks per reference or test serum are quantitatively harvested by trypsinization at day 7, then pooled and counted. Cells are subcultured by inoculating 2 105 WI-38 cells into new duplicate 25 cm2 flasks containing fresh growth medium that has been supplemented with the same lot of reference or test FBS used for the previous passage. The flasks are equilibrated and incubated as described in step 2. 4. The assay is continued as described through three passages, and the average number of cells/flask for each reference or test serum is calculated at the end of each passage. The relative growth rate (RGR) in each test serum during the final subculture is expressed as a fraction of the cell growth obtained in the reference serum: % RGR = average cells/flask in test serum average cells/flask in reference serum
Sf9 cell growth promotion assay. The insect assay assures that the FBS lot you purchase will support the growth of Sf9 cells. Upon receipt, all you have to do is heat-inactivate the serum for 30 min at 56C with mixing. This assay protocol can also be used for the biological performance assay of lactalbumin hydrolysate, yeastolate, and other growth supplements tested as raw materials. The accuracy of this assay is dependent on adherence to a routine cell maintenance schedule. Cell viability should be at least 80% as determined by trypan blue dye exclusion. This assay is performed on lots of certified FBS (base cat. nos. 16000, and 10082) and on selected lots of qualified FBS (base cat. nos. 26140 and 10099).
Sf9 cell growth assay methodology

1. Complete test media are prepared using a previously approved lot of Graces Insect Culture Medium supplemented with 10% test or reference heat-inactivated FBS. 2. Stock Sf9 cells are dispensed into 50 ml centrifuge tubes and centri fuged at 50 g for 5 min. Each tube is resuspended with complete medium containing test or reference heat-inactivated FBS at 10% concentration. 3. After inverting the tubes several times, a 1 ml sample is removed from each tube. A viable cell count is performed using trypan blue dye exclusion. 4. If viability is 80%, the tube is inverted several times and a 0.25 ml sample is removed from the tube. A cell count is then performed using a Coulter counter. 5. Cells are inoculated into Erlenmeyer flasks at a final concentration of 2.75 105 cells/ml. After inoculation, another cell count is performed to assure a cell density of 2.5 to 3.0 105 cells/ml. 6. The flasks are incubated on a shaker apparatus at 27C 1C at 8090 rpm for 96 hours. 7. Two 0.25 ml samples are removed from each flask and counted on the Coulter counter. A sample is also removed for determination of cell viability. 8. The relative cell number (RCN) is determined by comparing the test cell density to the control cell density.
Reference
Martinez, E.C., Cady, D.J., Grefrath, P.L., and Jayme, D.W. (1985) In Vitro 21, 30A.
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Additional Tests: Certified (US)

The following additional tests are performed on base cat. nos. 16000 and 10082. You will receive a Certificate of Analysis documenting the results of each quality control assay. Bacteriophage tests.2 Testing for the presence of bacteriophages using a plaque-assay method.3 A representative sample from each serum lot is added to tryptose phosphate agar containing a suspension of bacteriophage-sensitive E. coli (C-3000 or K-12). The mixture is then layered onto tryptose phosphate agar plates and incubated at 36C 2C for approximately 24 hours, and the plates are observed for plaque formation.
Performance Tests: Other Sera

Bovine sera and Newborn Calf sera. Each lot of Bovine Serum and Newborn Calf Serum (see Other Sera Ordering Information, pages 126128) is tested for its ability to support the growth of Vero cells over three subcultures. At each passage, the cells are subcultured to the original cell-inoculation density. Results are compared to parallel results obtained using control growth medium containing a previously characterized reference serum. Horse serum. Each lot of Horse Serum (see Other Sera Quality Control Tests, pages 132133) is tested for its ability to support the growth of Sp2/O-Ag14 (murine myeloma) cells in control medium containing the test lot of serum. Results are compared to parallel results obtained using control growth medium containing a previously characterized reference serum.
Biochemical Profile Tests

alkaline phosphatase bicarbonate bilirubin (total) blood urea nitrogen (BUN) BUN/creatinine ratio calcium chloride cholesterol creatinine gamma glutamyl transpeptidase (GGTP) glucose high-density lipoprotein (HDL) lactate dehydrogenase (LDH) low-density lipoprotein (LDL) potassium phosphorous (inorganic) serum glutamyl oxaloacetate transaminase (SGOT) sodium total iron binding capacity transferrin triglycerides (TG) uric acid
Cytotoxicity Tests: Other Sera

The performance testing procedures described in the preceding section are, by design, stringent tests for cytotoxicity as well. For some sera that do not undergo performance testing, we test specifically for cytotoxicity. These products are listed in the table below. Test samples are examined for the ability to support the attachment and proliferation of three adherent cell lines. The test and reference cultures are monitored for evidence of nutritional deficiency, abnormal morphology, or cytotoxicity.
Sera Tested for Cytotoxicity

Product Base cat. no. 16110 16070 26250 Chicken Serum
Hormone Profile Tests

estradiol insulin progesterone testosterone thyroxine (T4)
Lamb Serum Porcine Serum
Hemoglobin Test
All lots of US Certified FBS are guaranteed to be 15 mg/dl hemoglobin with 70% comprised of oxyhemoglobin.
Additional Tests: ES Cell-Qualified FBS

Gibco ES Cell-Qualified FBS is specially tested for the ability to sustain the undifferentiated cellular morphology of embryonic stem cells.
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Special Processing of Sera

Dialysed Sera For researchers requiring dialysed sera, we provide sera that are dialysed against 0.15 M NaCl using 10,000 molecular weight cut-off membrane until the glucose level is <5.0 mg/dl, as determined by the Glucose oxidase/peroxidase (GOD/POD) method (1, 2). The tangential flow filtration (TFF) process allows serum to be processed and final filtered on the same day. To prevent precipitation and inactivation of serum peptides, we do not perform exhaustive dialysis. Thus, we do not certify that all low molecular weight dialyzable components, such as amino acids, are totally removed by our processing. Heat Inactivated Sera We heat inactivate serum by mixing it in a water bath at a thermostatically controlled temperature of + 56o C for 30 mins. Ultra Low lgG FBS The advent of the Biotechnology industry and the production of biologicals from Cell Culture has highlighted some disadvantages in using Foetal Bovine Serum in certain production regimes. For example, the presence of immunoglobulins (in particular IgG) can cause difficulties in the purification of certain biologicals (e.g. Monoclonal Antibodies). We have developed a proprietary chromatographic procedure for the removal of gamma globulin from FBS. This procedure allows us to produce FBS with very low concentrations of IgG (<5mg/ml) while maintaining full biological activity in the sera. Afurther significant feature of serum produced using this process is the extremely low BVD antibody titre (Serum Index < 1). Charcoal Stripped FBS For researchers who require lower levels of hormones, especially estradiol, we offer this special FBS that has been processed using a proprietary charcoal/dextran treatment to reduce hormone and growth factor levels.
References
1. Keston, A. (abstract) 129th meeting of ACS, September 1956, 31C 2. Trinder, P (1969) Ann. Clin. Biochem. 6.24
Gamma Irradiated Serum Serum is one of the most commonly used supplements in many cell culture systems and as any biological derived product, it carries the risk of being contaminated by adventitious agents, particularly viruses and mycoplasma. Although mycoplasma can be successfully removed by filtration through 0.1 * pore size filter, the same is not true for most viruses. Various methods for eliminating virus contamination have been applied to serum. Gamma irradiation is the traditional method of choice for inactivating virus in serum as it enables effective virus inactivation over a wide spectrum of virus types, unlike UV irradiation.
Virus inactivation under experimental conditions (minimum dose of - 25kGy)

Spiked serum before treatment (pfu/ml) Spiked serum after gamma irradiation < 0.1 < 0.1 < 0.1 < 0.1* < 0.1 Reduction index for total process > 8.7 x 105 > 1.6 x 106 > 2.0 x 106 > 2.0 x 106 > 1.8 x 106
Virus BVD IBRV P13 PPV RE03

* Titre expressed as TCID50 values
8.7 x 104 1.6 x 105 2.0 x 10

5
2.0 x 105* 1.8 x 10

5
200
Notes:
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Notes:
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Appendix I
Gibco Universal Bags . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204 Prefilled Wave Bag . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207 new Lab Armortm Beads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208 PD-Directtm Bioprocess Services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209 Cell Culture basics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210 new Supply Centre by Life Technologiestm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213 Formulas and Formula Weights for Commonly Used Medium Components . . . . . . . . . . . . . . . . . 216 Troubleshooting: Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218 Useful Numbers for Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219 Buffer Systems for Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219 Specialty Media and Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219 Serum-free Media and Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220 Hybridoma Culture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220 Chinese Hamster Ovary (CHO) Cell Culture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221 Human Embryonic Kidney (293) Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221 Mammalian Cell Culture for Virus Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 221 Insect Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222 Neuronal Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 222 Blood and Bone Marrow Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223 Stem Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223 Other Mammalian Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223 Adaptation of Cell Cultures to a Serum-free Medium. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 224 Adaptation of Cell Cultures to an Alternative Serum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 225 Adaptation of Cells from Monolayer to Suspension Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . 225 Stability of GlutaMAX-I vs. L-Glutamine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226 Cryopreservation of Mammalian Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227 Thawing of Cryopreserved Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227 Dissociation of Cells from Primary Tissue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 228 Dissociation of Cells from Culture Vessels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229 Decontaminating Cultures with Antibiotics and Antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . 231 Use of Antibiotics and Antimycotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231 Conversion Factors, Units, and Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 232 Genetic Code and Amino Acid Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233
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Appendix I Reference Guide
Gibco media and reagents available in flexible packaging

Specifically designed to increase productivity and reduce costs Conveniently prefilled in a variety of formulations and sizes Easily integrated with most systemsunique universal design provides multiple connection options For over 49 years, gibco has built a reputation for innovation and service, with a commit-ment to quality. Our focus on packaging is no exception and our engineers have worked closely with industry leaders to extend the range of liquid delivery systems. Gibco universal bags are designed with common tubing connections for easy and con-venient integration into all your media applications. You can order your media prefilled in a 5, 10, or 20 L universal bag (Figure 1) and choose from a variety of stocked formulations.If your project requires a media and bag not offered prefilled, you can contact a knowl-edgeable member of Invitrogens cell culture custom team to assist you with selecting the right configuration. Packaging options are offered in a consistent film ranging from 250 mL up to 1,000 L.
Superior-quality film
The gibco universal bag incorporates an animal originfree 9101 film with an ultra low-density polyethylene media contact layer (Figure 2). It has been fully qualified* and widely used in many cell culture applications. Consistent film across gibcos bag offering makes scale up easier, eliminating the need for multiple validations. All bag sizes, ranging from 250 mL to 1,000 L, are manu-factured using the 9101 film Extensive documentation for the 9101 film is available and includes: USP Class VI, leachables, extractables, gas permeability, and biological and physical properties Superior gas transmission rates improve product consistency (Figure 4 Transparent film allows clear and improved view of bag contents (Figure 3) Proven stability for pH and cell performance ensure product quality (Figure Y with 2 graphs to be provided).
Figure 1. Gibco universal bags have a flexible four-por t design offering versatility and convenience. (1) female-threaded luer; (2) blood spike; (3) male-threaded luer; (4) MPC quick connect.
Figure 2. 9101 film construction details. 1. 50 m Polyethylen 2. 10 m Adhesive layer 3. 20 m Ethyl Vinyl Alcohol (Gas barrier 4. 10 m Adhesive Layer5. 230 m Ultra low-density polyethylene (Contact layer).
Figure 3. New 9101 film (on left) provides superb clarity of contents
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Flexible, four-port design

The gibco line of flexible packaging allows you to utilize and integrate a single-use, dis-posable container with a variety of automated cell culture instruments, disposable bioreac-tor systems, and sterile connection processes without the need for costly customization. Gibco universal bags are compatible with four different mechanical connection methods (blood spike, male-threaded luer, female-threaded luer, and MPC quick connect (Figure 5); featuring: Versatile and closed media transfer system utilizing three preconnected C-Flex tub-ing transfer lines Variety of tubing sizes for flow rate flexibility (ID OD): 1/8 1/4 (3.2 mm x 6.4 mm) 3/16 5/16 (4.8 mm x 7.9 mm) 3/8 5/8 (9.5 mm x 15.9 mm) Compatibility with the most commonly used thermoplastic tubing
Improved packaging
Gibco universal bags are shipped in a reinforced, 100% recyclable container. Made from heavyweight corrugate material, these boxes provide a solid system to transport and store your gibco universal bags. Durable five-layer handles provide a strong hold to comfortably and confidently carry your media.
Figure 5. Four-port design. Gibco universal bags have a flexible four-port design offering versatility and convenience: (1) female-threaded luer, (2) blood spike, (3) male-threaded luer, and (4) MPC quick connect.
Figure 4. Gas transmission rates. The gibco universal bag shows superior gas transmission rates in compari-son to current bags on the market.
Figure 6. Gibco universal bags are ideal for many cell culture applications.
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Gibco Universal Bags

Order gibco media and reagents in universal bags from Life Technologiestm today.
Product Balanced salts DPBS without calcium chloride, magnesium chloride, or phenol red Quantity Cat. No.
5L 10 L 20 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 5L 10 L 20 L 5L 5L 5L 10 L 20 L 10 L 20 L 10 L 20 L 10 L 20 L
14190 -342 14190-359 14190-367 10010-002 10010-001 14170-195 14170-203 11965-167 11965-175 10566-032 10566-040 11995-115 11995-123 10569-069 10569-077 12430-112 12430-120 10564-037 10564-045 11330-099 11330-107 11875-168 11875-176 61870-143 61870-150 72400-146 72400-153 22400-147 22400-154 15230-253 15230-261 15230-279 12605-093 12604-054 10902-161 10902-179 10902-187 12658-035 12658-001 10743-001 10743-002 12338-001 12338-002
PBS without phenol red, pH 7.4 HBSS without calcium chloride, magnesium chloride, or magnesium sulfate Basal media DMEM high glucose with L-glutamine, without sodium pyruvate DMEM high glucose with GlutaMAX-I, without sodium pyruvate DMEM high glucose, with L-glutamine and sodium pyruvate DMEM high glucose, with GlutaMAX-I and sodium pyruvate DMEM with L-glutamine and HEPES buffer, without sodium pyruvate DMEM with GlutaMAX-I and HEPES buffer, without sodium pyruvate DMEM/F-12 with L-glutamine and HEPES buffer RPMI Medium 1640 with L-glutamine RPMI Medium 1640 with GlutaMAX-I RPMI Medium 1640 with GlutaMAX(TM)-I and HEPES buffer RPMI Medium 1640 with L-glutamine and HEPES buffer Reagents Distilled Water
TrypLE Express 1X with phenol red TrypLE Express 1X without phenol red Expression Media Sf-900 II with L-glutamine
Sf-900 III with L-glutamine CD CHO FreeStyle 293
For additional information, to place your order, or to request a custom format, formulation, or film construction, please contact Life Technologiestm Customer Service or visit www.invitrogen.com/universalbags. Make to stock product.
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Prefilled Wave Bag

The Cellbag media bag from Wave Biotech is now available prefilled with gibco media Ready to shipmedia formulations include Sf-900 II and FreeStyle 293 ConvenientCellbag chambers are filled and ready to use Customizableyou choose the gibco medium formulation and volume
Appendix I
Product Sf-900 II SFM FreeStyle 293 Expression Medium Quantity 10 L fill in 20 L Wave Cellbag 7 L fill in 20 L Wave Cellbag 8 L fill in 20 L Wave Cellbag Cat. no. 0070037BA 0070049BA 0070049BB
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Lab Armor Beads

Replace the water in your lab bath to help prevent contamination
Standing, warm water in your laboratory water bath can introduce unknown bacteria, yeasts, fungi, mycoplasmas, and chemical contaminants into your lab. These contaminants can affect your cell culture reagents, resulting in distorted data and unpredictable outcomes. Lab Armor Beads replace the water in your laboratory water bath and reduce the potential for contamination of your media, reagents, and cells. With Lab Armor Beads, you can protect your research and use your lab bath with confidence.
Lab Armor Beads are low-maintenance and highly consistent

Cleanwater baths must be routinely monitored, cleaned, and maintained; Lab Armor Beads only need a light spray of biodegradable disinfectant every 24 weeks Convenientno need to plan around warm-up times, or worry about burnout when water evaporates OrganizedLab Armor Beads naturally hold items in place, no matter the size or shape; racks, floats, and weights are not required Consistentgreater temperature uniformity and maintenance than water
Confidence and reproducibility with Lab Armor Beads and gibco products
Gibco cell culture products deliver reproducible results and awardwinning service while helping you reduce the risk of contamination in your lab. Use gibco cell culture products and Lab Armor Beads together every time you perform cell culture. Remember to include Lab Armor Beads in your next order of gibco cell culture products. Gibco cell culture products and Lab Armor Beads help you achieve reproducible results every day. Learn more at www.invitrogen.com/labarmorbeads.
Lab Armor Beads are the environmentally responsible choice

Eliminates biocidesliquid disinfectants used for water bath maintenance are harmful to the environment; periodic use of a biodegradable disinfectant like ethanol is all that is required to keep Lab Armor Beads free of contamination Recyclableformed from ultrapure, recyclable material Energy-efficientset at 35C, Lab Armor Beads consume less than one half of the energy required by water
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PD-Direct Bioprocess Services

Invitrogen has over 49 years of experience in the cell culture business and over this time, the Life Technologiestm name and reputation has stood as the recognized leader in cell culture systems. Life Technologiestm expanded on this tradition in innovative cell culture products through the creation of a team focused on process development, PD-Direct Bioprocess Services. The PD-Direct team responds to core customer needs by offering customers comprehensive service from cell line and medium development to process optimization (Figure 1). Our cell line services include: Parental cell line optimization through natural adaptation or accelerated evolution utilizing the Revolution technology Preoptimized, serum-free FreeStyle expression services to produce a protein within a week in suspension HEK or 293 cultures Cell line engineering with proprietar y molecular biology technologies Recombinant cell line creation with CHO cell lines that incorporates industry-leading and IP-friendly vector components Clone selection or enrichment through the use of the ClonePixFL high- throughput fluorescent screening technology Expression enhancement with the Revolution technology for difficultto-produce proteins
Appendix I
Upstream Optimization Services

The PD-Direct team brings together the best scientists and technologies in the area of upstream optimization. Through a proven approach of applying high-throughput technologies to process design within an integrated workflow, our systematic approach yields performance improvements in your manufacturing process. The PD-Direct upstream services range from clone-specific process and media optimization to platform development bridging Cell line development, cloning, basal media, and feed supplements. Our ability to bring this step change in performance comes from different elements: Unrivaled breadth of historical experience in media screening and media formulation across cell lines and cell types A high-throughput workflow integrating multi-factor statistical experimental design with Simcell miniturised bioreactors and Hamilton Starplus automated media formulation. Comprehensive media analytics to provide vital in-process information Component harmonization and elimination of unnecessary or inhibitory components to reduce system complexity Expertise from cloning through large-scale production that empowers integrated media platforms that minimize adaptation time and effort
Get your process going

Through a partnership with Invitrogen, you can join industry leaders who have discovered the difference that Invitrogens innovation can bring to their projects. Whether a point solution for upstream optimization or cell line creation, or an integrated process development solution, PD-Direct Bioprocess Services is the trusted partner that brings the expertise and breadth of Invitrogen to solving your process development needs. Get your process going by visiting us at www.invitrogen.com/pddirect or emailing us at pd.direct@invitrogen.com.
Cell Line Services

The science of cell line development has become complex. Increasing production demands with shorter development timelines require a breadth of investments in platforms, technologies, and personnel. Factors such as transfection efficiency, cell productivity, media conditions, and cell line stability must be incorporated into an overall program of cell line optimization. From a myriad of expression enhancers to high-throughput screening, the challenges of optimization continue to grow. Additionally, intellectual property, technology harmonization, and final product quality are other considerations that complicate the process. In response, Life Technologiestm has integrated an array of services to simplify cell line development, through PD-Direct Bioprocess Services. We offer you a suite of the best cell line development technologies from across the industry and our in-depth knowledge. This unique combination will yield a high-quality production cell line that will serve as the foundation for your therapeutic product development for years to come.
Figure 1PD-Direct Bioprocess Services expands your process development options.
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Cell Culture basics

This section provides information on the fundamentals of cell culture, including the selection of the appropriate cell line for your experiments, media requirements for cell culture, adherent versus suspension culture, and morphologies of continuous cell lines available from Life Technologiestm. Note that the following information is an introduction to the basics of cell culture, and it is intended as a starting point in your investigations. For more in-depth information, we recommend that you consult published literature and books, as well as the manuals and product information sheets provided with the products you are using.
Acquiring Cell Lines

You may establish your own culture from primary cells, or you may choose to buy established cell cultures from commercial or non-profit suppliers (i.e., cell banks). Reputable suppliers provide high quality cell lines that are carefully tested for their integrity and to ensure that the culture is free from contaminants. We advise against borrowing cultures from other laboratories because they carry a high risk of contamination. Regardless of their source, make sure that all new cell lines are tested for mycoplasma contamination before you begin to use them. Life Technologiestm offers a variety of primary cultures and established cell lines, reagents, media, sera, and growth factors for your cell culture experiments. The Appendix section contains a list of the more commonly used cell lines available from Life Technologiestm. For more information on Life Technologiestm and gibco products, refer to www.invitrogen.com. One of the major advantages of cell culture is the ability to manipulate the physicochemical (i.e., temperature, pH, osmotic pressure, O2 and CO2 tension) and the physiological environment (i.e., hormone and nutrient concentrations) in which the cells propagate. With the exception of temperature, the culture environment is controlled by the growth media. While the physiological environment of the culture is not as well defined as its physicochemical environment, a better understanding of the components of serum, the identification of the growth factors necessary for proliferation, and a better appreciation of the microenvironment of cells in culture (i.e., cell-cell interactions, diffusion of gases, interactions with the matrix) now allow the culture of certain cell lines in serum-free media.
Cell Lines
Selecting the Appropriate Cell Line
Consider the following criteria for selecting the appropriate cell line for your experiments Species: Non-human and non-primate cell lines usually have fewer biosafety restrictions, but ultimately your experiments will dictate whether to use speciesspecific cultures or not. Functional characteristics: What is the purpose of your experiments? For example, liver- and kidney-derived cell lines may be more suitable for toxicity testing. Finite or continuous: While choosing from finite cell lines may give you more options to express the correct functions, continuous cell lines are often easier to clone and maintain. Normal or transformed: Transformed cell lines usually have an increased growth rate and higher plating efficiency, are continuous, and require less serum in media, but they have undergone a permanent change in their phenotype through a genetic transformation. Growth conditions and characteristics: What are your requirements with respect to growth rate, saturation density, cloning efficiency, and the ability to grow in suspension? For example, to express a recombinant protein in high yields, you might want to choose a cell line with a fast growth rate and an ability to grow in suspension. Other criteria: If you are using a finite cell line, are there sufficient stocks available? Is the cell line well-characterized, or do you have to perform the validation yourself? If you are using an abnormal cell line, do you have an equivalent normal cell line that you can use as a control? Is the cell line stable? If not, how easy it is to clone it and generate sufficient frozen stocks for your experiments?
Adherent vs Suspension Culture

There are two basic systems for growing cells in culture, as monolayers on an artificial substrate (i.e., adherent culture) or free-floating in the culture medium (suspension culture). The majority of the cells derived from vertebrates, with the exception of hematopoietic cell lines and a few others, are anchorage-dependent and have to be cultured on a suitable substrate that is specifically treated to allow cell adhesion and spreading (i.e., tissue-culture treated). However, many cell lines can also be adapted for suspension culture. Similarly, most of the commercially available insect cell lines grow well in monolayer or suspension culture. Cells that are cultured in suspension can be maintained in culture flasks that are not tissue-culture treated, but as the culture volume to surface area is increased beyond which adequate gas exchange is hindered (usually 0.20.5 mL/cm2), the medium requires agitation. This agitation is usually achieved with a magnetic stirrer or rotating spinner flasks.
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Adherent Culture Appropriate for most cell types, including primary cultures. Requires periodic passaging, but allows easy visual inspection under inverted microscope. Cells are dissociated enzymatically (e.g., TrypLE Express, trypsin) or mechanically. Growth is limited by surface area, which may limit product yields. Requires tissue-culture treated vessel. Used for cytology, harvesting products continuously, and many research applications.
Suspension Culture Appropriate for cells adapted to suspension culture and a few other cell lines that are nonadhesive (e.g., hematopoietic). Easier to passage, but requires daily cell counts and viability determination to follow growth patterns; culture can be diluted to stimulate growth. Does not require enzymatic or mechanical dissociation. Growth is limited by concentration of cells in the medium, which allows easy scale-up. Can be maintained in culture vessels that are not tissue-culture treated, but requires agitation (i.e., shaking or stirring) for adequate gas exchange. Used for bulk protein production, batch harvesting, and many research applications.
Appendix I
Media
The culture medium is the most important component of the culture environment, because it provides the necessary nutrients, growth factors, and hormones for cell growth, as well as regulating the pH and the osmotic pressure of the culture. Although initial cell culture experiments were performed using natural media obtained from tissue extracts and body fluids, the need for standardization, media quality, and increased demand led to the development of defined media. The three basic classes of media are basal media, reduced-serum media, and serum-free media, which differ in their requirement for supplementation with serum. Serum is vitally important as a source of growth and adhesion factors, hormones, lipids and minerals for the culture of cells in basal media. In addition, serum also regulates cell membrane permeability and serves as a carrier for lipids, enzymes, micronutrients, and trace elements into the cell. However, using serum in media has a number of disadvantages including high cost, problems with standardization, specificity, variability, and unwanted effects such as stimulation or inhibition of growth and/or cellular function on certain cell cultures. If the serum is not obtained from reputable source, contamination can also pose a serious threat to successful cell culture experiments. To address this threat, all Invitrogen and gobco products, including sera, are tested for contamination and guaranteed for their quality, safety, consistency, and regulatory compliance. Basal Media The majority of cell lines grow well in basal media, which contain amino acids, vitamins, inorganic salts, and a carbon source such as glucose, but these basal media formulations must be further supplemented with serum. Reduced-Serum Media Another strategy to reduce the undesire d effects of serum in cell culture experiments is to use reduced-serum media. Reduced-serum media are basal media formulations enriched with nutrients and animal-derived factors, which reduce the amount of serum that is needed.
Serum-Free Media Serum-free media (SFM) circumvents issues with using animal sera by replacing the serum with appropriate nutritional and hormonal formulations. Serum-free media formulations exist for many primary cultures and cell lines, including recombinant protein producing lines of Chinese Hamster Ovary (CHO), various hybridoma cell lines, the insect lines Sf9 and Sf21 (Spodoptera frugiperda), and for cell lines that act as hosts for viral production (e.g., 293, VERO, MDCK, MDBK), and others. One of the major advantages of using serum-free media is the ability to make the medium selective for specific cell types by choosing the appropriate combination of growth factors. The table below lists the advantages and disadvantages of serum-free media.
Advantages Increased definition More consistent performance Easier purification and downstream processing Precise evaluation of cellular functions Increased productivity Better control over physiological response Enhanced detection of cellular mediators Disadvantages Requirement for cell type-specific media formulations Need for higher degree of reagent purity Slower growth
Invitrogen offers a wide range of classical basal media, reducedserum media, and serumfree media, as well as sera, growth factors, supplements, antibiotics, and reagents for your cell culture experiments. The Appendix section contains a list of the more commonly used cell culture products available from Life Technologiestm. For more information on Life Technologies tm and gibco cell culture products, refer to www.invitrogen.com.
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pH
Most normal mammalian cell lines grow well at pH 7.4, and there is very little variability among different cell strains. However, some transformed cell lines have been shown to grow better at slightly more acidic environments (pH 7.07.4), and some normal fibroblast cell lines prefer slightly more basic environments (pH 7.47.7). Insect cell lines such as Sf9 and Sf21 grow optimally at pH 6.2.
Temperature
The optimal temperature for cell culture largely depends on the body temperature of the host from which the cells were isolated, and to a lesser degree on the anatomical variation in temperature (e.g., temperature of the skin may be lower than the temperature of skeletal muscle). Overheating is a more serious problem than underheating for cell cultures; therefore, often the temperature in the incubator is set slightly lower than the optimal temperature. Most human and mammalian cell lines are maintained at 36C to 37C for optimal growth. Insect cells are cultured at 27C for optimal growth; they grow more slowly at lower temperatures and at temperatures between 27C and 30C. Above 30C, the viability of insect cells decreases, and the cells do not recover even after they are returned to 27C. Avian cell lines require 38.5C for maximum growth. Although these cells can also be maintained at 37C, they will grow more slowly. Cell lines derived from cold-blooded animals (e.g., amphibians, coldwater fish) tolerate a wide temperature range between 15C and 26C. Note that cell culture conditions vary for each cell type. The consequences of deviating from the culture conditions required for a particular cell type can range from the expression of aberrant phenotypes to a complete failure of the cell culture. We therefore recommend that you familiarize yourself with your cell line of interest, and closely follow the instructions provided with each product you are using in your experiments.
CO2
The growth medium controls the pH of the culture and buffers the cells in culture against changes in the pH. Usually, this buffering is achieved by including an organic (e.g., HEPES) or CO2-bicarbonate based buffer. Because the pH of the medium is dependent on the delicate balance of dissolved carbon dioxide (CO2) and bicarbonate (HCO3), changes in the atmospheric CO2 can alter the pH of the medium. Therefore, it is necessary to use exogenous CO2 when using media buffered with a CO2bicarbonate based buffer, especially if the cells are cultured in open dishes or transformed cell lines are cultured at high concentrations. While most researchers usually use 57% CO2 in air, 410% CO2 is common for most cell culture experiments. However, each medium has a recommended CO2 tension and bicarbonate concentration to achieve the correct pH and osmolality; refer to the media manufacturers instructions for more information.
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Maximise your research time with 24/7 access to a wide variety of brands
24/7 access means never having to run out of essential products at that crucial stage in your experiments. Just a phonecall or email to the dedicated Supply Centre team results in queries being handled efficiently and always in a friendly manner.
Lizzie Gault University of Glasgow, UK
Appendix I
Supply Centre by Life Technologies is an electronic, customisable onsite stocking programme offering a premium consignment delivery service.
Right product in the right place at the right time Easy to use Customised to your needs Reduced transaction cost Dedicated support
Contact supplycentre.query@lifetech.com for more information.
www.invitrogen.com/supplycentre
With the pressure to publish and difficulty to acquire funding for academic research, Supply Centre by Life technologies has saved us a great amount of cost and administration time. Working without the Supply Centre is no longer an option.
Dr. Bart Vanaudenaerde KU Leuven, Belgium
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Introducing Supply Centre by Life Technologies
Supply Centre by Life Technologies. Its what you need. Now.

Life Technologiestm is committed to creating an environment in which the biomedical sciences flourish. Our efforts are focused on a simple goal; how to better meet the needs of scientists. To this end, we have designed our Supply Centre programme for the efficient and convenient storage of products at our customers sites, because researchers should not have to put their work on hold while a critical enzyme, reagent, or kit is being shipped. Convenience Instant access to staple research products Total Workflow Solution - Customise product mix to best suit your Research requirements Cost effective Reduce transaction costs, consolidate shipping and streamline invoice processing Flexibility Ability to order non-stock products Zero risk All appliances and stock owned & maintained by Life Technologies
The Supply Centre Management System (SCMS)

The Supply Centre Management System is a purpose built, web-based interface that allows you to run your Supply Centre easily and efficiently. This flexible system is designed to support your existing business rules, to integrate with your existing procurement platform, and to simplify ordering. The Supply Centre Management System allows full visibility of order history, current stock list, account specific pricing and provides access to administrative reports.
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A fully customised solution

Each installation is fully customised to suit the customers individual requirements with regard to: Product Mix - Select from the broad portfolio of Life Technologies brands. Location - May be located in either a Lab, Stores area or public area Appliance(s) - Choice of appliance based on product mix and/or selected site or location. Delivery options - Weekly, Bi-Weekly or Monthly, delivery day selected by the customer Invoicing Options - Choice of Standard or Consolidated invoicing Integration with existing procurement solutions
A premium service with premium support

Our commitment to providing you with 24/7 access to the products you need does not end with on-site stocking. From installation to invoicing, your dedicated Supply Centre team is available to assist with any query you may have. Why wait? To find out more about how to install a Supply Centre at your site, please contact your local Life Technologies sales representative or contact the Supply Centre team at: supplycentre.query@lifetech.com
Appendix I
Seamless integration with your existing procurement system

We recognise that many customers have procurement workflows within their organisations and to support eBusiness and efficiency, purchasing is often carried out within these environments. To support these customer requirements Life Technologiestm can offer a full suite of integration capabilities including: B2B integration to SCMS (Supply Centre Management System) Punchout (OCI/Roundtripping) B2B integration to catalogue-based systems These options are provided at no cost to our customers through our dedicated eBusiness team who can initiate and manage the project at each stage of the integration. With over 150 live B2B projects and over 400 Supply Centres installed in Europe, Life Technologiestm has the experience to meet your integration needs.
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Formulas and Formula Weights for Commonly Used Medium Components

Formula weight (to nearest whole number) 1,255 404 267 347 605 347 89 103 80 1,236 174 211 176 132 150 133 152 244 397 111 147 218 236 477 387 253 140 461 238 768 371 170 250 121 176 240 313 243 251 227 264 267 134 Formula weight (to nearest whole number) 397 404 278 786 786 441 512 180 116 180 216 180 176 217 203 147 146 329 307 75 188 283 238 155 210 131 136 159 180 131 583 146 131 280 206 146 183 120 246 95 203 134 342
Compound Actinomycin D5H2O Adenine sulfate2H2O Adenosine Adenosine 5-phosphate Adenosine 5-triphosphate disodium salt3H2O Adenylic acid L-Alanine L--Amino-n-butyric acid Ammonium nitrate Ammonium molybdate4H2O L-Arginine L-Arginine hydrochloride Ascorbic acid L-Asparagine L-AsparagineH2O L-Aspartic acid 8-Azaguanine Biotin Calciferol (Vitamin D2) Calcium chloride Calcium chloride2H2O Calcium lactate Calcium nitrate4H2O D-Calcium pantothenate Cholesterol Choline bitartrate Choline chloride Co-carboxylase Cobalt chloride6H2O Coenzyme A Colcemid * Cupric chloride2H2O Cupric sulfate5H2O L-Cysteine L-Cysteine hydrochlorideH2O L-Cystine L-Cystine dihydrochloride Cytidine 2-Deoxyadenosine 2-Deoxycytidine 2-Deoxycytidine hydrochloride 2-Deoxyguanosine Deoxyribose
Formula C62H86N12O165H2O (C5H5N5)2H2SO42H2O C10H13N5O4 C10H14N5O7P C10H14N5Na2O13P33H2O C10H14N5O7P C3H7NO2 C4H9NO2 NH4NO3 (NH4)6Mo7O244H2O C6H14N4O2 C6H14N4O2HCl C6H8O6 C4H8N2O3 C4H8N2O3H2O C4H7NO4 C4H4N6O C10H16N2O3S C28H44O CaCl2 CaCl22H2O C6H10CaO6 Ca(NO3)24H2O C18H32CaN2O10 C27H46O C9H19NO7 C5H14ClNO C12H19ClN4O7P2S CoCl26H2O C21H36N7O16P3S C21H25NO5 CuCl22H2O CuSO45H2O C3H7NO2S C3H7NO2SHClH2O C6H12N2O4S2 C6H12N2O4S22HCl C9H13N3O5 C10H13N5O3 C9H13N3O4 C9H13N3O4HCl C10H13N5O4 C10H5O4
Compound Ergocalciferol (Vitamin D2) Ferric nitrate9H2O Ferrous sulfate7H2O Flavine FAD (flavin adenine dinucleotide) Folic acid Folinic acid, calcium salt D-Fructose Fumaric acid D(+) Galactose D-Glucosamine hydrochloride D-Glucose D-Glucuronolactone GlutaMAX-I GlutaMAX-II L-Glutamic acid L-Glutamine Glutathione sodium salt Glutathione (reduced) Glycine Guanine hydrochloride Guanosine HEPES L-Histidine L-Histidine hydrochlorideH2O L-Hydroxyproline Hypoxanthine Hypoxanthine sodium salt i-Inositol L-Isoleucine Kanamycin sulfate -Ketoglutaric acid L-Leucine Linoleic acid Lipoic acid L-Lysine L-Lysine hydrochloride Magnesium sulfate Magnesium sulfate7H2O Magnesium chloride Magnesium chloride6H2O Malic acid Maltose
Formula C28H44O Fe(NO3)39H2O FeSO47H2O C27H33N9O15P2 C27H33N9O15P2 C19H19N7O6 C20H21CaN7O7 C6H12O6 C4H4O4 C6H12O6 C6H13NO5HCl C6H12O6 C6H8O6 C8H15N3O4 C7H13N3O4 C5H9NO4 C5H10N2O3 C10H16N3O6SNa C10H17N3O6S C2H5NO2 C5H5N5OHCl C10H13N5O5 C8H18N2O4S C6H9N3O2 C6H9N3O2HClH2O C5H9NO3 C5H4N4O C5H4N4ONa C6H12O6 C6H13NO2 C18H36N4O11H2SO4 C5H6O5 C6H13NO2 C18H32O2 C8H14O2S2 C6H14N2O2 C6H14N2O2HCl MgSO4 MgSO47H2O MgCl2 MgCl26H2O C4H6O5 C12H12O11
* Colcemid is a trademark of Ciba Limited.
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Compound Manganese chloride4H2O Manganese sulfateH2O Menadione (Vitamin K3) Menadione sodium bisulfate L-Methionine 5-Methylcytosine 5-Methyl-deoxycytidine Methyl linoleate NAD (nicotinamide adenine dinucleotide) NADP (nicotinamide adenine dinucleotide phosphate) Niacinamide Nicotinic acid L-Ornithine hydrochloride Para-aminobenzoic acid Phenol red Potassium chloride Potassium nitrate Potassium phosphate monobasic L-Phenylalanine L-Proline Putrescine dihydrochloride Pyridoxal hydrochloride Pyridoxal 5-phosphate Pyridoxine hydrochloride Riboflavin Riboflavin 5-phosphate sodium salt D-Ribose L-Serine Sodium acetate Sodium acetate3H2O Sodium bicarbonate Sodium chloride Sodium glucuronateH2O
Formula MnCl24H2O MnSO4H2O C11H8O2 C11H9NaO5S C5H11NO2S C5H7N3O C10H15N3O3 C19H34O2 C21H27N7O14P2 C21H28N7O17P3 C6H6N2O C6H5NO2 C5H13ClN2O2 C7H7NO2 C19H12O5SNa2 KCl KNO3 KH2PO4 C9H11NO2 C5H9NO2 C4H14Cl2N2 C8H9NO3HCl C8H10NO6P C8H11NO3HCl C17H20N4O6 C17H20N4NaO9P C5H10O5 C3H7NO3 C2H3O2Na C2H3O2Na3H2O NaHCO3 NaCl C6H11NaO7H2O
Formula weight (to nearest whole number) 198 169 172 276 149 125 225 295 663 743 122 123 169 137 398 75 101 136 165 115 161 204 247 206 376 478 150 105 82 136 84 58 236
Compound Sodium phosphate dibasic Sodium phosphate dibasic7H2O Sodium phosphate monobasicH2O Sodium pyruvate Sodium selenite Sodium succinate6H2O Succinic acid Sucrose L-Taurine Thiamine hydrochloride Thiamine monophosphate L-Threonine Thymidine Thymine (5-Methyluracil) DL--Tocopherol acetate DL--Tocopherol phosphate sodium salt Trehalose2H2O L-Tryptophan L-Tyrosine L-Tyrosine disodium salt Uracil Uridine Uridine 5-triphosphate Uridine 5-triphosphate trisodium salt2H2O L-Valine Vitamin A Vitamin A (acetate) Vitamin B12 Xanthine Xanthine sodium salt Zinc chloride Zinc sulfate7H2O
Formula Na2HPO4 Na2HPO47H2O NaH2PO4H2O C3H3O3Na Na2SeO3 C4H4O4Na26H2O C4H6O4 C12H22O11 C2H7NO3S C12H17ClN4OSHCl C12H20N4O8P2S C4H9NO3 C10H14N2O5 C5H6N2O2 C31H52O3 C29H55O9P2Na42H2O C12H22O112H2O C11H12N2O2 C9H11NO3 C9H9NO32Na C4H4N2O2 C9H12N2O6 C9H15N2O15P3 C9H12N2O15P3Na32H2O C5H11NO2 C20H30O C22H32O2 C63H88CoN14O14P C5H4N4O2 C5H3NaN4O2 ZnCl2 ZnSO47H2O
Formula weight (to nearest whole number) 142 268 138 110 173 270 118 342 125 337 442 119 242 126 473 738 378 204 181 225 112 244 484 586 117 286 328 1,355 152 174 136 288
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Troubleshooting: Cell Culture*

Problem Rapid pH shift in medium Possible cause Incorrect carbon dioxide (CO2) tension Suggested solution Increase or decrease percentage of CO2 in the incubator based on concentration of sodium bicarbonate in medium. For sodium bicarbonate concentrations of 2.0 to 3.7 g/L, use CO2 amounts of 5% to 10%, respectively. Switch to CO2-Independent Medium (Cat. no. 18045-088). Loosen caps one-quarter turn. Add HEPES buffer to a final concentration of 10 to 25 mM. Use an Earles saltsbased medium in a CO2, environment and a Hanks saltsbased medium in atmospheric conditions. Discard culture and medium. Try to decontaminate culture (see Decontaminating Cultures with Antibiotics and Antimycotics, page 207). Rinse glassware in deionized, distilled water several times, then sterilize.
Overly tight caps on tissue culture flasks Insufficient bicarbonate buffering Incorrect salts in medium Bacterial, yeast, or fungal contamination Precipitate in medium, no change in pH Residual phosphate left over from detergent washing, which can precipitate powdered medium components Frozen medium Precipitate in medium, change in pH Cells not adhering to culture vessel Bacterial or fungal contamination Overly trypsinized cells Mycoplasma contamination No attachment factors in medium Suspension cells clumping together Presence of calcium and magnesium ions Mycoplasma contamination Cell lysis and release of DNA resulting from overdigestion with proteolytic enzymes Primary cell culture is contaminated Decreased growth of culture Contamination of primary tissue carried over to culture Change in medium or serum
Warm medium to 37C and swirl to dissolve. If precipitate remains, discard medium. Discard medium. Try to decontaminate culture (see Decontaminating Cultures with Antibiotics and Antimycotics, page 207). Trypsinize for a shorter time, or use less trypsin (see Dissociation of Cells from Culture Vessels, page 205). Segregate culture and test for mycoplasma infection. Clean hood and incubator. If culture is contaminated, discard. For serum-free formulations, be sure they contain attachment factors or use coated plates. Wash cells in a balanced salt solution without calcium and magnesium. Gently pipet cells to obtain a single-cell suspension. Segregate culture and test for mycoplasma infection. Clean hood and incubator. If culture is contaminated, discard. Treat cells with 0.001% DNase I. Rinse cells with PBS and seed fresh medium. Wash tissue pieces several times in a balanced salt solution containing a higher concentration of antibiotics and antimycotics before culturing. Compare medium formulations for differences in glucose, amino acids, and other components. Compare the old lot of serum with the new lot in a growth experiment. Increase initial cell inoculum. Adapt cells sequentially to the new medium. Remove medium and add fresh medium.
Depletion, absence, or breakdown of essential growth-promoting components such as L-glutamine or growth factors
Low-level bacterial or fungal contamination Improper storage of reagents
Low initial cell inoculum Senescence of finite culture Mycoplasma contamination Death of culture No CO2 in the incubator Temperature fluctuations in the incubator Use of Fungizone Reagent or antibiotics at toxic concentrations Cell damage during thawing or cryopreservation Incorrect osmotic pressure in medium
Supplement medium with growth-promoting components. Substitute GlutaMAX-I (Cat. no. 35050-061) for L-glutamine in the medium. Grow culture without antibiotics. If contaminated, discard culture. Try to decontaminate, (see Decontaminating Cultures with Antibiotics and Antimycotics, page 207). Store sera at 5C to 20C. Store media at 2C to 8C in the dark. Store complete media at 2C to 8C in the dark and use within 2 weeks. Minimize exposure of sera and media to light. Increase viable cell inoculum. Discard culture; obtain new cell stock. Segregate culture and test for mycoplasma infection. Clean hood and incubator. If culture is contaminated, discard. Monitor rate of CO2 use in incubators to determine when to change tanks. Check line connections frequently for leaks. Avoid opening and closing incubator doors. Monitor temperature of incubator. Use less Fungizone Reagent (see Use of Antibiotics and Antimycotics, page 207 for appropriate concentrations). Note: For serum-free media reduce antibiotics 10 times. Obtain another aliquot of cells (see Cryopreservation of Mammalian Cells, page 203). Check osmolality of complete medium. Note: Most mammalian cells can tolerate an osmolality of 260 to 350 mOsm/kg. Addition of reagents such as HEPES and drugs may affect osmolality. Note: Insect cells require media with a higher osmolality (340 to 380 mOsm/kg) than mammalian cells. Remove medium and add fresh medium.
Buildup of toxic metabolites in the medium
* There are several common problems encountered when culturing cells. Problems in primary cultures may have different causes than the same problem in established cell lines. Below are some of the common problems encountered when culturing cells, along with their possible causes and suggested solutions. Fungizone is a trademark of E.R. Squibb & Sons, LLC.
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Useful Numbers for Cell Culture*

Surface area (mm2) Dishes 35 mm 60 mm 100 mm 150 mm Cluster plates 6-well 12-well 24-well Flasks T-25 T-75 T-160 2,500 7,500 16,000 0.7 106 2.1 10
6
Seeding density 0.3 106 0.8 106 2.2 106 5.0 106 0.3 106 0.1 10
6
Cells at confluency 1.2 106 3.2 106 8.8 106 20.0 106 1.2 106 0.4 10
6
Versene (ml of 0.53 mM EDTA) 1 3 5 10 2 1 0.5 3 5 10
Trypsin (ml of 0.05% trypsin, 0.53 mM EDTA) 1 2 3 8 2 1 0.5 3 5 10
Growth medium (ml)
Appendix I
962 2,827 7,854 17,671 962 401 200
2 3 10 20 35 12 0.51.0 35 815 1530
0.05 106
0.2 106 2.8 106 8.4 106 18.4 106
4.6 106
* There are various sizes of dishes and flasks used for cell culture. Some useful numbers such as surface area and volumes of dissociation solutions are given below for various sizes of culture vessels. The number of cells on a confluent plate, dish, or flask will vary with cell type. For this table, HeLa cells were used.
Buffer Systems for Cell Culture

Buffered salt solutions for cell culture can be divided into two principal types: those designed to equilibrate with atmospheric conditions, and those designed to equilibrate with a gas phase containing 5% to 10% CO2. Media utilizing Hanks salts are designed for atmospheric equilibration, based upon the physiologically relevant pK a of phosphoric acid. These media contain bicarbonate ions for carboxylation and similar biological processes.1 Use of Hanks salts in a CO2 incubator will result in a rapid drop in pH of the culture medium. This will cause the phenol red indicator to turn yellow. Media based on Earles salts are buffered with a bicarbonate/carbonic acid system and rely upon the physiologically relevant pK a for carbonic acid and the equilibration of gaseous and liquid carbon dioxide to maintain the pH in a CO2-equilibrated incubator.1 Use of Earles salts in atmospheric conditions will result in a rapid rise in pH of the culture medium. This will cause the phenol red indicator to turn purple.
Reference
1. Jayme, D.W. and Blackman, K.E. (1985) Adv. in Biotechnol. Processes 5, 9.
Specialty Media and Applications*

Cell type A wide range of cells routinely cultured in serum-supplemented medium Specialty media Advanced DMEM and Advanced MEM Opti-MEM I Reduced-Serum Medium Hybridomas Amniocytes, chorionic villus cells with ethanolamine hydrochloride AmnioMAX-II Complete Medium AmnioMAX-C100 Complete Medium (basal medium and supplement) Lymphocytes Bone marrow and peripheral blood PB-MAX Karyotyping Medium MarrowMAX Bone Marrow Medium Application Enhanced standard basal formulations which allow for a 90% reduction in the required serum supplementation (no cell adaptation needed) A modified Eagles MEM medium intended to reduce the amount of serum supplementation by 50% Permits some hybridomas to grow serum-free In vitro propagation of primary human amniotic fluid cells for karyotyping In vitro propagation of primary human amniotic fluid cells for karyotyping. Supplement contains growth supplements (serum) and antibiotics Fully supplemented (including FBS) RPMI-based medium for short-term cultivation of peripheral blood lymphocytes A fully supplemented medium developed specifically to support bone marrow and peripheral blood cell culture for in vitro cytogenetic analysis of hematological disease
* Specialty media are media supplied with or supplemented with serum designed for specific applications.
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Serum-free Media and Applications

Serum-free media are designed to grow a specific cell type or perform a specific application in the absence of serum. Serum-free media (SFM) represent an important tool that allows cell culture to be done with a defined set of conditions as free as possible of confounding variables. Advantages of using serum-free media: Increased definition More consistent performance Easier purification and downstream processing Precise evaluations of cellular function Increased growth and/or productivity Better control over physiological responsiveness Enhanced detection of cellular mediators Certain applications may require the addition of growth factors and/or cytokines. Note: Customized cell culture media are available. Submit your requests to our Custom Products Services group for timely evaluation and production of the formulation and delivery system that best fits your needs.
Glossary
Serum-free media gibco Serum-free Media do not require supplementation with serum, but may contain discrete proteins or bulk protein fractions. Protein-free mediagibco Protein-free Media contain no proteins, but may contain plant or yeast hydrolysates. Many are animal originfree. Chemically defined mediagibco Chemically Defined Media contain no proteins, hydrolysates, or components of unknown composition. These media are animal originfree and all components have a known chemical structure. Animal originfree productsgibco animal originfree products do not contain material directly derived from animal tissues, cells, or body fluids of higher eukaryotic organisms, such as mammals (including humans), fish, birds, insects, etc. The term animal origin does not pertain to lower eukaryotic organisms such as the higher plants, fungi, protozoa, and algae, nor does it include prokaryotic organisms such as bacteria or blue-green algae.
Hybridoma Culture
Product
Optimized for CD Hybridoma Medium* CD Hybridoma AGT Dry granular format of CD Hybridoma Medium Hybridoma-SFM Low-protein 20 g/ml Human, mouse, rat hybridomas, myelomas, NS0, NS-1, and other steroid-dependent cells when used with 250X Cholesterol Lipid Concentrate
Application Growth and MAb production Can be used to express other proteins in engineered myeloma cell lines
Quantity 1,000 ml 11L 1 10 L 500 ml 1,000 ml 1,000 ml
Cat. no. 11279-023 12372-025 12372-017 12045-084 12045-076 12040-077
Human, mouse, rat hybridomas, myelomas
Growth and MAb production Can be used to express other proteins in engineered myeloma cell lines Growth and MAb production Can be used to express other proteins in engineered myeloma cell lines
Human, mouse, rat hybridomas, myelomas PFHM-II Protein-free Hybridoma Medium
* Drug Master File available.
220
Chinese Hamster Ovary (CHO) Cell Culture

Product
Optimized for CD OptiCHO Medium CD CHO Medium* CD CHO Medium AGT * Dry granular format of CD CHO Medium Adherent CHO cells Suspension CHO cells (including CHO-S cells) Suspension CHO cells (including CHO-S cells) Adherent CHO cells Suspension CHO cells (including CHO-S cells) Suspension CHO cells (including CHO-S cells)
Application Growth and production of recombinant proteins Growth and production of recombinant proteins
Quantity 1,000 ml 6 1,000 ml 500 ml 1,000 ml 11L 1 10 L 500 ml
Cat. no. 12681-011 12681-029
Appendix I
10743-011 10743-029 12490-017 12490-025 097-0182DJ 12052-114 12052-098 096-0334DJ 097-0147DK 11619-012
CD CHO-A Medium
CHO-S-SFM II* Available without hypoxanthine and thymidine (Cat. no. 31033-020)
Growth and production of recombinant proteins in suspension culture Growth and production of recombinant proteins in suspension culture Growth and production of recombinant proteins in adherent culture
500 ml 1,000 ml 500 ml 1,000 ml 1.5 ml
CHO III PFM* CHO III-A-PFM
CHO-S Cells, adapted to CD CHO Medium, CHO-S-SFM II and CHO III PFM
Human Embryonic Kidney (293) Cell Culture

Product
N IMA A FREE L O RIGIN N IMA A FREE L O RIGIN
Optimized for CD 293 Medium* CD 293 AGT * Dry granular format of CD 293 Medium Suspension 293 cells and HeLa S3 cells Suspension 293-F cells Suspension 293 cells (including 293-F, 293-H)
Application Growth and recombinant protein or adenovirus production in suspension culture
Quantity 1,000 ml 11L 1 10 L 1,000 ml
Cat. no. 11913-019 12529-020 12529-012 11686-029
293 SFM II* Low-protein <10 g/ml
Growth and recombinant protein or adenovirus production in suspension culture Growth and transfection in suspension culture
1,000 ml 6 1,000 ml 8 L fill in 20 L Wave Cellbag
12338-018 12338-026 0070049BB 11625-019 11631-017 R790-07
293-F Cells, adapted to CD 293 Medium and 293 SFM II 293-H Cells, adapted to CD 293 Medium and 293 SFM II FreeStyle 293-F Cells, adapted to FreeStyle 293 Expression Medium
1.5 ml 1.5 ml 1 107 cells
Mammalian Cell Culture for Virus Production

Product
Optimized for VP-SFM* Low-protein <10 g/ml VP-SFM AGT * Dry granular format of VP-SFM OptiPRO SFM* Low protein <10 g/ml Protein Expression Medium (PEM) Adherent MDCK, VERO, PK-15, MDBK, BHK-21, and related cells used in virus production Recombinant proteins in PER.C6 cells Suspension BHK-21 cells, adherent VERO, COS-7L, MDCK, HEp-2
Application For culture of kidney epithelial and related cells used in virus production
Quantity 1,000 ml 11L 1 10 L
Cat. no. 11681-020 12559-027 12559-019 12309-019 12661-013
For culture of kidney epithelial and related cells used in virus production Supports growth of parental PER.C6, HEK293, and HeLa cells in suspension culture
1,000 ml 1,000 ml
* Drug Master File available. PER.C6 is a trademark of Crucell Holland B.V.
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Insect Cell Culture

Product Express Five SFM Optimized for Suspension BTI-TN-5B1-4 insect cells Application Growth and maintenance of cells used for the baculovirus expression vector system (BEVS) for adherent or suspension culture Large-scale production of recombinant protein expressed by BEVS Sf-900 II SFM* Suspension Sf9, Sf21 (Spodoptera frugiperda), TN368 cells (Trichoplusia ni) Growth and maintenance of cells used for BEVS for adherent or suspension culture Large-scale production of recombinant protein expressed by BEVS 500 ml 1,000 ml 6 1,000 ml (case) 5L 10 L 20 L 10 L fill in 20 L Wave Cellbag 1.5 ml 3 10 cells/ml 1.5 ml 1.5 ml
6
Quantity 1,000 ml
Cat. no. 10486-025
10902-096 10902-088 10902-104 10902-161 10902-070 10902-187 0070037BA 10831-014 B855-02 11496-015 11497-013
D.Mel-2 Cells High Five Cells, adapted to Express Five SFM Sf9 Cells, adapted to Sf-900 II SFM Sf21 Cells, adapted to Sf-900 II SFM
Neuronal Cell Culture

Product Neurobasal Medium Neurobasal Medium without phenol red Neurobasal-A Medium Neurobasal-A Medium without phenol red B-27 Serum-free Supplement B-27 Supplement minus AO B-27 Supplement minus Vitamin A (Retinoic acid) N-2 Supplement Primary embryonic hippocampal neurons, tumor cell lines of neuronal origin (PC12, B104, N1E-115 and NS20) Primary glial cells, tumor cell lines of glial origin (U-251, MGsp, C62BD, RN-22), astrocytes, microglia, oligodendrocytes Adult and postnatal neurons (>1 week old) Primary embryonic hippocampal neurons; primary neurons from striatum, substantia nigra, septum Growth and maintenance. Minimizes glial cell proliferation B-27 minus AO to study free-radical damage, apoptosis (B-27 minus AO is formulated without any cortical antioxidants) Study growth of CNS progenitor or stem cells Maintenance of primary neurons (low protein, <125 g/ml) Growth and maintenance of neuronal tumor cell lines Growth and maintenance of primary and serial tumor glial cells 1 ml 17503-012 Optimized for Fetal neurons Application Basal medium lacking excitatory amino acids used in conjunction with supplements to make a complete serum-free medium Long-term growth of neurons Quantity Cat. no. 500 ml 500 ml 500 ml 500 ml 10 ml 10 ml 10 ml 5 ml 21103-049 12348-017 10888-022 12349-015 17504-044 10889-038 12587-010 17502-048
G-5 Supplement
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Blood and Bone Marrow Culture

Product AIM V Medium, liquid,* research grade, with HSA AIM V Medium, liquid research grade, with BSA AIM V Medium, liquid, therapeutic grade Macrophage-SFM Macrophages, monocytes Optimized for Lymphocytes, macrophages, monocytes, lymphoid cell lines Lymphocytes, macrophages, monocytes, and lymphoid cell lines Application Research use only, growth of TIL cells, CTL cells, and monocytes Research use only, growth of TIL cells, CTL cells, and monocytes Growth of tumor infiltrating lymphocytes (TIL cells), cytotoxic T cells, monocytes, and ex vivo activation Growth and maintenance (addition of GM-CSF may be necessary). Demonstration of macrophage phagocytosis. Activation of cells to kill tumor cells with -interferon or lipopolysaccharide supplementation Growth and maintenance of human hematopoietic progenitor cells (addition of factors necessary). Study synergistic/individual effects of growth Growth of bone marrow, peripheral blood, and hematopoietic cells in short-term in vitro culture Quantity 500 ml 1,000 ml 500 ml 1,000 ml 10 L 500 ml Cat. no. 12055-091 12055-083
Appendix I
31035-025 087-0112DK 087-0112BK 12065-074
StemPro-34 SFM Supplied with Hematopoietic StemPro Nutrient Supplement MarrowMAX Bone Marrow Medium
* Drug Master File available
Human hematopoietic progenitor cells (CD34+) from bone marrow, peripheral blood, or neonatal cord blood Bone marrow, peripheral blood, and hematopoietic cells
500 ml
10639-011
100 ml
12260-014
Stem Cell Culture

Product KnocKout DMEM KnocKout Serum Replacement StemPro hESC SFM MesenPRO RS Medium Growth and expansion of human embryonic stem cells (hESCs) Growth of human mesenchymal stem cells (MSCs) in culture Optimized for Murine and human embryonic stem (ES) cells Application Growth and maintenance of undifferentiated ES cells for production of transgenic mice. Growth and maintenance of both human and murine ES cells used in differentiation studies Maintains pluripotency in a growing list of hESC lines including BG01, BG02, BG03, H1, H9, and BG01V Improvement of MSC expansion. Expansion of cells for multiple passages while maintaining multipotential phenotype (i.e., differentiation into osteogenic, chondrogenic, and adipogenic lineages) Quantity 500 ml 500 ml 1 kit 1 kit Cat. no. 10829-018 10828-028 A1000701 12746-012
Other Mammalian Cell Culture

Product Human Endothelial-SFM* Optimized for Primary and secondary human umbilical venous, microvascular, and arterial endothelial cells Primary human, monkey, and rat hepatocytes Human epidermal keratinocytes and cervical epithelial cells (will not support fibroblast or melanocyte cells) Application Growth and maintenance to study cell-cell interactions, injury analysis, atherosclerosis, signal transduction, cytokine production, and cell matrix interaction. Requires supplementation with bFGF, EGF, and fibronectin (sold separately) Maintenance of hepatocytes (cytochrome P450 induction maintained >9 days) Growth and maintenance for dermal substitutes, gene therapy, and in vitro toxicology Without bovine pituitary Human epidermal keratinocytes cervical epithelial cells (will not support fibroblast or melanocyte cells) Low-protein (<25 g/ml) without BPE medium for cultivation of keratinocytes. Can be used for cervical epithelial cells for studies involving the human papilloma virus Quantity 500 ml Cat. no. 11111-044
Hepatozyme-SFM Keratinocyte-SFM (with EGF, BPE)*
500 ml 500 ml
17705-021 17005-042
Keratinocyte-SFM (without CaCl2)* Defined Keratinocyte-SFM*
500 ml 500 ml
37010-022 10744-019
Primary Human Keratinocytes, adapted to Defined Keratinocyte-SFM

1 ml
12332-011
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Adaptation of Cell Cultures to a Serum-free Medium

There are two approaches to adapt cells to serum-free medium (SFM): 1. Direct adaptationcells switched from serum-supplemented medium into SFM. 2. Sequential adaptation or weaningcells switched from serumsupplemented medium into SFM in several steps. (Sequential adaptation tends to be less harsh on cells than direct adaptation.) To adapt cells to SFM, it is critical that the culture is: In mid-logarithmic phase >90% viable Seeded at a higher initial cell inoculum during adaptation Below are general protocols for adaptation. Initial cell inocula, cell yields, and culture periods are intended as starting points for subsequent optimization.
Antibiotics
It is best not to use antibiotics in serum-free media. If you do, we recommend that you use 5- to 10-fold less than you would in a serumsupplemented medium. This is because serum proteins tend to bind a certain amount of the antibiotic added; without these serum proteins the level of antibiotic may be toxic to certain cells.
Higher Density
Cells must be in the mid-logarithmic phase of growth with viability >90% prior to adaptation. Sequential adaptation may be necessary. Seeding cultures at a higher density than normal at each passage during SFM adaptation may help the process. Because some percentage of cells may not survive in the new culture environment, having more cells present will increase the number of viable cells to further passage.
Direct Adaptation
Some cells can be directly adapted from serum-containing medium to SFM. For direct adaptation, the cell inoculum should be 2.5 105 to 3.5 105 cells/ml. Subculture cells when the cell density is 1 106 to 3 106 cells/ml. Cells are fully adapted to SFM when the cell density is 2 106 to 4 106 cells/ml after 4 to 7 days in culture. Stock cultures of cells adapted to SFM should be subcultured in SFM every 3 to 5 days when the cell density is 1 106 to 3 106 with 90% viability.
Clumping
Cell clumping often occurs during adaptation to SFM. We recommend that you gently triturate the clumps to break them up when passaging cells.
Morphology
It is not uncommon to see slight changes in cellular morphology during and after adaptation to SFM. As long as doubling times and viability remain good, slight changes in morphology should not be a reason for concern.
Sequential Adaptation
Sequential adaptation is Invitrogens preferred method for adapting cells to serum-free media (SFM), with a typical conversion being:
Passage 1 Passage 2 Passage 3 Passage 4 75% serum-supplemented medium: 25% SFM 50% serum-supplemented medium: 50% SFM 25% serum-supplemented medium: 75% SFM 100% SFM
Adaptation with Conditioned Medium

An alternate method for adaptation to SFM involves the use of conditioned medium. This is medium the cells have been growing in for one full passage. If you choose this method, you can facilitate adaptation as follows:
Passage 1 Passage 2 Passage 3 Passage 4 Passage 5 100% serum-supplemented medium 50% medium from passage 1: 50% SFM 50% medium from passage 2: 50% SFM 50% medium from passage 3: 50% SFM 100% SFM
Because the change from 75% to 100% SFM may be too stressful for your cells, you may need to carry the cells for 23 passages in a 10% serum-supplemented medium: 90% SFM mixture. Most cell lines can be considered fully adapted after 3 passages in 100% SFM. Occasionally you may have trouble getting your cells past a certain step even before going 100% SFM. If this happens, go back and passage the cells 23 times in the previous ratio of serum-supplemented medium to serum-free medium.
Points to Consider in Serum-free Culture

Overall, cells in serum-free culture are more sensitive to extremes of pH, temperature, osmolality, mechanical forces, and enzyme treatment.
Whichever adaptation method you choose, we strongly recommend that you always take these precautions: Make a frozen stock of the cells in the serum-supplemented medium prior to adaptation. Keep a culture going of the cells in each prior condition when starting the next level of adaptation as a fallback if the cells do not survive in the next passage.
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Adaptation of Cell Cultures to an Alternative Serum

Many cells adapt best with a sequential method of adaptation. Culture cells with a 1:1 (v:v) blend of the current medium with FBS and the medium with the alternative serum. Reduce the amount of current medium over successive passages by blending the media as follows: 1:2, 1:4, 1:16, and 100% medium with alternative. Passage cells 2 or 3 times with each change in serum composition for adaptation. Cultures may be switched to the alternative serum directly from FBS. Initially, use the alternative serum at the same concentration as the FBS. A lag in growth may occur. Allow 2 to 3 passages for the growth rate to return to its previous level.
Appendix I
Adaptation of Cells from Monolayer to Suspension Culture

Adaptation of adherent cells to suspension culture may be required to grow cells to a higher density or for the growth of cells in serum-free medium. This protocol describes the procedure for cells grown in T-25 or T-75 flasks. Approximately 6 to 8 T-75 flasks provide enough HeLa cells (5 107) to seed a suspension culture in a spinner or shake flask. Note: The number of starting flasks will vary depending on the cell line. For best results remember the following: Use cells 50% to 80% confluent Use cells with >90% viability 1. Remove growth medium. 2. Wash cells using 1X D-PBS without calcium and magnesium (3 ml for T-25 flask or 5 ml for T-75 flask). Discard the wash solution. 3. Add trypsin-EDTA (0.05% trypsin, 0.53 mM EDTA4Na; 3 ml for T-25 flask or 5 ml for T-75 flask) to the side of the flask opposite the cells. Briefly swirl the flask. Discard all but enough trypsin-EDTA to leave a thin film of liquid over the cells. Incubate the cells at 37C for 5 to 10 min. Monitor the cells during this incubation under a microscope. When they round up, tap the flask to dislodge the cells. 4. Add growth medium (6 ml for T-25 flask or 10 ml for a T-75 flask) to the cell suspension. Note: For serum-free medium, add soybean trypsin inhibitor. Usually, using 1:1 (v:v) 0.25 mg/ml trypsin inhibitor solution to trypsin will inhibit trypsin. 5. Transfer the cell suspension to a 15 ml centrifuge tube. Centrifuge at 100 g for 4 min. Remove the supernatant (trypsin-EDTA mixed with growth medium) by aspiration. 6. Resuspend the cells in 5 ml of growth medium. Centrifuge at 100 g for 4 min. Remove the supernatant by aspiration. Note: For serum-free medium resuspend the cells in D-PBS without calcium or magnesium. 7. Resuspend the cells in 5 ml of growth medium. Count the number of viable cells. 8. Dilute cells in growth medium to obtain a density of 5 105 cells/ml. 9. Subculture in a sterile spinner or shake flask. Leave sufficient headspace for adequate gas exchange (100 ml of medium in a 250 ml spinner flask or 75 to 100 ml of medium in a 250 ml shake flask). Note: For serum-free medium that does not contain a shear protectant, add Pluronic F-68 to a final concentration of 0.1%. 10. Loosen caps for gas exchange and place at 37C in a humidified atmosphere of 5% to 10% CO2 in air. For spinner flasks, use an impeller speed of 75 to 95 rpm. For shake flasks, rotate on a shaker platform at 125 to 135 rpm. 11. Check viable cell density daily to establish growth kinetics in suspension culture. 12. When the cell density is 1 106 cells/ml (or on day 4 post-planting) passage the cells at a density of 5 105 cells/ml. Note: Centrifugation and the addition of fresh growth medium may be necessary if the cell density has not reached 1 106 cells/ml by day 4. 13. Once cell density reaches 1 106 cells/ml with 90% viability by day 3 post-planting for 3 passages, the cells are considered adapted to suspension culture. Seeding density can be reduced to 2 105 to 3 105 cells/ml.
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Stability of GlutaMAX-I vs. L-Glutamine

Glutamine is an essential nutrient in cell cultures for energy production as well as protein and nucleic acid synthesis. A drawback to glutamine in cell culture media is spontaneous breakdown once in solution. Glutamine breakdown generates ammonia as a by-product, which is toxic to the cells 1 and can affect protein glycosylation.2,3 One strategy to minimize generation of ammonia is to replace the L-glutamine in media with the glutamine dipeptide, GlutaMAX-I. Figures 1 and 2 demonstrate the improved stability of the glutamine dipeptide and reduction of spontaneous ammonia generation. An additional benefit to using GlutaMAX-I in place of L-glutamine in cell culture is the extension of culture time, potentially reducing the number of times the cells must be passaged. Figure 3 is a comparison of MDBK cells cultured in DMEM with 10% fetal bovine serum (FBS) and either glutamine or GlutaMAX-I. Cells cultured in GlutaMAX-I reach peak density two days later, and viability declines less rapidly than in cultures with glutamine supplementation. Many widely used gibco media formulations are available with the GlutaMAX dipeptide substituted for L-glutamine. See page 24 for a complete listing of products. Additionally, the GlutaMAX dipeptide is available as a 200 mM solution and can be substituted for glutamine on an equimolar basis. It can be used with most cell lines with equivalent performance and substituted in both serum-supplemented and serum-free formulations requiring L-glutamine. The slight increase of the lag phase is attributed to the time needed to release the peptidase and digest the dipeptide. This allows a gradual increase in availability of L-glutamine to the cells.1
Common Cell Lines Cultured with GlutaMAX-I

MDBK MDCK BHK CHO C-33 A HT-29 293 Per.C6 * BG01v H1 HepG2 HELA AE-1 3D9 Ns0/1 Sp2/0 D3 J1 Jurkat Swiss 3T3 Bovine kidney Canine kidney Hamster kidney Hamster ovary Human cervical cancer Human colon cancer Human embryonic kidney Human embryonic retinoblastoma Human ES cell Human ES cell Human hepatoblastoma used in AlgiMatrix Human ovary Mouse hybridoma Mouse hybridoma Myeloma Myeloma Mouse ES cell lines Mouse ES cell lines Human T cell leukemia Swiss mouse fibroblast
References
1. Hassell, T., Gleave, S., and Butler, M. (1991) Growth Inhibition in Cell Culture. Applied Biochemistry and Biotechnology, v. 30, pp 3041. 2. Yang, M., and Butler, M. (2002) Effects on Ammonia and Glucosamine on the Heterogeneity of Erythropoietin Glycoforms. Biotechnology Progress, v. 18, 129138. 3. Yang, M., and Butler, M. (2000) Effect of Ammonia on the Glycosylation of Human Recombinant Erythropoietin in Culture. Biotechnology Progress, v. 16, 751759.
* PER.C6 is a trademark of Crucell Holland B.V.
1,000 900
70
10 9
Total cell count ( 105)
60
GlutaMAX-I
Ammonia (mg/L)
800 700
8 7 6 5 4 3 2 1 0
GlutaMAX-I L-glutamine
50 40 30 20 10 0
L-glutamine
mg/L
600 500 400 300 200 0 1 2 3 4 5 6 7

GlutaMAX-I L-glutamine
Day
Day
Day
Figure 1Stability of GlutaMAX-I vs. L-glutamine in DMEM at 37C. DMEM was supplemented with GlutaMAX-I or L-glutamine, aliquoted into vials, and stored at 37C. Samples were taken daily and frozen at 20C. Levels of GlutaMAX-I and L-glutamine were determined by HPLC.
Figure 2Ammonia levels in supplemented media stored at 37C. DMEM was supplemented with GlutaMAX-I or L-glutamine, aliquoted into vials, and stored at 37C. Samples were taken daily and frozen at 20C. Levels of ammonia were determined by HPLC.
Figure 3Growth of MDBK cells in DMEM supplemented with L-glutamine or GlutaMAX-I and 10% FBS. MDBK cells were seeded at approximately 1 105 cells/flask in DMEM with 10% FBS and L-glutamine or GlutaMAX-I in 25 cm2 T-flasks.
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Cryopreservation of Mammalian Cells

Mammalian cells are cryopreserved to avoid loss by contamination, to minimize genetic change in continuous cell lines, and to avoid aging and transformation in finite cell lines. Before cryopreserving, cells should be characterized and checked for contamination. Recovery Cell Culture Freezing Medium (Cat. no. 12648-010) is a complete cryopreservation medium for mammalian cell cultures. With Recovery Cell Culture Freezing Medium, you will increase cell viability an average of 25% and save time without measuring and mixing as required to prepare your own cryoprotectant. Recovery Cell Culture Freezing Medium is prepared in DMEM (high glucose) with FBS and calf serum and 10% DMSO. There are several common media used to freeze cells. For serumcontaining medium, the constituents may be as follows: Complete medium containing 10% glycerol Complete medium containing 10% (dimethylsulfoxide) (DMSO) 50% cell-conditioned medium with 50% fresh medium with 10% glycerol 50% cell-conditioned medium with 50% fresh medium with 10% DMSO For serum-free medium, some of the common medium constituents may be: 50% cell-conditioned, serum-free medium with 50% fresh serum-free medium containing 7.5% DMSO fresh, serum-free medium containing 7.5% DMSO and 10% cell culturegrade BSA 3. Centrifuge at ~200 g for 5 min to pellet cells. Using a pipette, withdraw the supernatant down to the smallest volume without disturbing the cells. 4. Resuspend the cells in freezing medium at 5 10 6 to 1 10 7 cells/ml. 5. Aliquot into cryogenic storage vials. Place vials on wet ice or in a 4C refrigerator, and start the freezing procedure within 5 min. 6. Cells are frozen slowly at 1C/min. This can be done by programmable coolers or by placing vials in an insulated box in a 70C to 90C freezer, then transferring to liquid nitrogen storage.1
Appendix I
Reference
1. Freshney, R. (1987) Culture of Animal Cells: A Manual of Basic Technique, p. 220, Alan R. Liss, Inc., New York.
Thawing of Cryopreserved Cells

Cr yopreser ved cells are fragile and require gentle handling. Cryopreserved cells are thawed quickly and plated directly into complete growth medium. If cells are particularly sensitive to cryopreservative (DMSO or glycerol), they are centrifuged to remove cryopreservative and then plated into complete growth medium. The following are suggested procedures for thawing cryopreserved cells.
Suspension Cultures
1. Count the number of viable cells to be cryopreserved. Cells should be in log phase. Centrifuge the cells at ~200 to 400 g for 5 min to pellet cells. Using a pipette, remove the supernatant down to the smallest volume without disturbing the cells. 2. Resuspend cells in freezing medium at 1 107 to 5 107 cells/ml for serum-containing medium, or 0.5 107 to 1 107 cells/ml for serumfree medium. 3. Aliquot into cryogenic storage vials. Place vials on wet ice or in a 4C refrigerator, and start the freezing procedure within 5 min. 4. Cells are frozen slowly at 1C/min. This can be done by programmable coolers or by placing vials in an insulated box in a 70C to 90C freezer, then transferring to liquid nitrogen storage.1
Direct Plating Method

1. Remove cells from storage and thaw quickly in a 37C water bath. 2. Plate cells directly with complete growth medium. Use 10 to 20 ml of complete growth medium per 1 ml of frozen cells. Perform a viable cell count. Cell inoculum should be at least 3 105 viable cells/ml. 3. Culture cells for 12 to 24 hr. Replace medium with fresh complete growth medium to remove the cryopreservative.
Centrifugation Method
1. Remove cells from storage and thaw quickly in a 37C water bath. 2. Place 1 to 2 ml of frozen cells in ~25 ml of complete growth medium. Mix very gently. 3. Centrifuge the cells at ~80 g for 2 to 3 min. 4. Discard supernatant. 5. Gently resuspend the cells in complete growth medium and perform a viable cell count. 6. Plate the cells. Cell inoculum should be at least 3 105 viable cells/ml.
Adherent Cultures
1. Detach cells from the substrate with dissociating agents. Detach as gently as possible to minimize damage to the cells (see Dissociation of Cells from Culture Vessels, page 204). 2. Resuspend the detached cells in complete growth medium and establish the viable cell count.
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Dissociation of Cells from Primary Tissue

A common method to obtain single-cell suspensions from primary tissue is enzymatic disaggregation. Expose the cells to enzymes for a minimal amount of time to preserve maximum viability. The following procedures disaggregate whole tissue to obtain a high yield of viable cells.1 3. Incubate at 37C for 4 to 18 hr. Addition of 3 mM CaCl2 increases the efficiency of dissociation. 4. Filter the cell suspension through a sterile stainless steel or nylon mesh to separate the dispersed cells and tissue fragments from the larger pieces. Fresh collagenase can be added to the fragments if further disaggregation is required. 5. Wash the suspension several times by centrifugation in HBSS. 6. Resuspend the pellet in culture medium. Count and seed the cells for culture.
Trypsin
1. After dissecting off unusable tissue, mince the remaining tissue into 3 to 4 mm pieces with a sterile scalpel or scissors. Wash the tissue pieces by resuspending in a balanced salt solution without calcium and magnesium. Allow the tissue pieces to settle, and remove the supernatant. Repeat the wash 2 or 3 times. 2. Place the container with the tissue pieces on ice, and remove any remaining supernatant. Add 0.25% trypsin in a balanced salt solution without calcium or magnesium (~1 ml of trypsin for every 100 mg of tissue). 3. Incubate at 4C for 6 to 18 hr to maximize penetration of the enzyme with little trypsin activity. 4. Decant and discard the trypsin from the tissue pieces. Incubate the tissue pieces with residual trypsin at 37C for 20 to 30 min. 5. Add warm complete medium to the tissue pieces and gently disperse the tissue by pipetting. If using a serum-free medium, also add soybean trypsin inhibitor. 6. Filter the cell suspension through sterile stainless steel mesh (100 to 200 m) to completely disperse any remaining tissue. Count and seed the cells for culture.
Dispase
1. Mince tissue into 3 to 4 mm pieces with a sterile scalpel or scissors. Wash the tissue pieces several times in a calcium- and magnesiumfree balanced salt solution. 2. Add dispase (0.6 to 2.4 units/ml in calcium- and magnesium-free balanced salt solution). 3. Incubate at 37C for 20 min to several hours. 4. Filter the cell suspension through sterile stainless steel or nylon mesh to separate the dispersed cells and tissue fragments from the larger pieces. Fresh dispase can be added to the fragments if further disaggregation is required. 5. Wash the suspension several times by centrifugation in the balanced salt solution. 6. Resuspend the pellet in culture medium. Count and seed the cells for culture.
Collagenase
1. Mince tissue into 3 to 4 mm pieces with a sterile scalpel or scissors. Wash the tissue pieces several times with Hanks Balanced Salt Solution (HBSS). 2. Add collagenase (50 to 200 units/ml in HBSS).
Reference
1. Freshney, R. (1987) Culture of Animal Cells: A Manual of Basic Technique, p. 117, Alan R. Liss, Inc., New York.
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Dissociation of Cells from Culture Vessels

The following is a general procedure to rapidly remove various cell lines from the substratum while maintaining cellular integrity. This procedure is not meant to be universally applicable for all cell lines. The optimal conditions and concentrations employed for individual systems should be determined empirically. Monitor cell viability at the time of subculturing Cell viability should be greater than 90% For serum-free media, it is advisable to decrease the amount of trypsin used 1. Remove and discard the spent cell culture medium. 2. Wash the cells using a balanced salt solution without calcium and magnesium or wash with EDTA (see table on next page to determine the correct wash solution). Add the wash solution to the side of the flask opposite the cells. Rinse the cell sheet by rocking the flask for 1 to 2 min and discard wash solution. 3. Add the dissociation solution of choice (Table 1, next page) at 2 to 3 ml/25 cm2 to the side of the flask opposite the cells. Be sure that the dissociation solution covers the cell sheet. Incubate the flask at 37C. Rock the flask gently. Generally, cells are dissociated in 5 to 15 min. The time needed to dissociate cells will vary according to cell line. Monitor the process carefully to avoid cell damage. Flasks of cell lines that are difficult to remove from the substratum may be tapped to expedite removal. 4. When the cells are completely detached, stand the flask in the upright position to allow the cells to drain to the bottom of the flask. Add complete medium to the flask. Disperse the cells by pipetting repeatedly over the surface of the monolayer. Count and subculture the cells. Note: For serum-free media, add soybean trypsin inhibitor. Usually, using 1:1 (v:v) 0.25 mg/ml trypsin inhibitor solution to trypsin will inhibit trypsin.
TrypLE Products
TrypLE products are based on a recombinant enzyme that replaces animal trypsin for the dissociation of adherent mammalian cells from plastic. It can be stored refrigerated and is stable at room temperature for six months. TrypLE Express works on a wide range of cells and requires little or no protocol changes. Inactivation with trypsin inhibitors is not required. 1. Decant the medium from the flask. Rinse the flask with 5 ml of Dulbeccos Phosphate Buffered Saline (D-PBS) without calcium or magnesium (base cat. no. 14190). Decant the D-PBS. 2. Add an appropriate volume (i.e., 2 ml in a 75 cm2 flask) of prewarmed TrypLE to the flask. Rock the vessel to coat the cell sheet completely. 3. Incubate at 37C until the cells have detached (observe at 5 minute intervals). Gently rap the vessel to dislodge the cells. 4. Dilute in 2 to 5 ml of cell culture growth medium and transfer the cell suspension to a 15 ml centrifuge tube. 5. Centrifuge for 5 to 10 minutes at 100 g. Discard the supernatant and suspend the cell pellet in 2 to 5 ml of fresh growth medium. 6. Determine the viable and total counts. 7. Seed the flasks according to normal protocol. 8. Incubate and subculture according to normal protocol depending on your cell type. Note: Use of soybean trypsin inhibitor is not recommended.
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Table 1Dissociation of cells from culture vessels.

Culture type Most continuous cell lines Strongly adherent continuous cell lines Many cells at early passage Continuous cell lines where integrity of cell surface proteins is important Wash solution HBSS or PBS without calcium and magnesium D-PBS HBSS or PBS without calcium and magnesium Dissociation solution Trypsin 0.25% in balanced salt solution without calcium and magnesium TrypLE products Trypsin 0.05% in 0.53 mM EDTA TrypLE products Weakly adherent epithelial cells Transformed fibroblasts Primary cells where integrity of cell surface proteins is required Strongly adherent early-passage cell lines Epithelial cells* Strongly adherent cells* Epithelial cells Some tumor cells Thick cultures, multiple layers* Dense cultures that are collagen rich All adherent cultures HBSS or PBS without calcium and magnesium 1 mM EDTA Dispase 0.6 to 2.4 units/ml in PBS without calcium and magnesium Trypsin 0.25% Collagenase 200 units/ml, 1 mM EDTA in balanced salt solution without calcium and magnesium Scrape off cell sheet HBSS or PBS without calcium and magnesium 0.5 mM to 1 mM EDTA 0.5 mM to 1 mM EDTA HBSS or PBS without calcium and magnesium EDTA, glycerol in sodium citrate Products 0.25% Trypsin (liquid) 2.5% Trypsin (liquid) (10X) Trypsin Powder (1:250) TrypLE Select (liquid) TrypLE Express (liquid) 0.05% Trypsin 0.53 mM EDTA (liquid) 0.5% Trypsin 5.3 mM EDTA (10X) (liquid) TrypLE Select (liquid) TrypLE Express (liquid) Enzyme-free Cell Dissociation Buffers Hanks based PBS based TrypLE Select (liquid) TrypLE Express (liquid) 0.25% Trypsin 1 mM EDTA (liquid) Trypsin Powder (1:250) Dispase Versene 1:5,000 (0.53 mM EDTA in PBS) Dispase 0.25% Trypsin 1 mM EDTA Versene 1:5,000 (0.53 mM EDTA in PBS) Trypsin Powder (1:250) Dispase 0.25% Trypsin 1 mM EDTA Collagenase 17100, 17101, 17102, 17104 Trypsin Powder (1:250) Cat. no. 15050-065 15090-046 27250-018 12563-011 12605-010 25300-054 15400-054 12563-011 12605-010 13150-016 13151-014 12563-011 12605-010 25200-056 27250-018 17105-041 15040-066 17105-041 25200-056 15040-066 27250-018 17105-041 25200-056 27250-018
TrypLE products Trypsin 0.25% in 1 mM EDTA Dispase 0.6 to 2.4 units/ml in PBS 0.5 mM to 1 mM EDTA Dispase 0.6 to 2.4 units/ml in PBS Trypsin 0.25% with 1 mM EDTA
* Some cells may be sensitive to EDTA. Scraping may cause mechanical damage and will not yield a single-cell suspension. Note: Adapted from Freshney, R. (1987) Culture of Animal Cells: A Manual of Basic Technique, p. 129, Alan R. Liss, Inc., New York.
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Decontaminating Cultures with Antibiotics and Antimycotics

When an irreplaceable culture becomes contaminated, researchers may attempt to eliminate or control the contamination. First, determine if the contamination is bacteria, fungus, mycoplasma, or yeast. Isolate the contaminated culture from other cell lines. Clean incubators and laminar flow hoods with a laboratory disinfectant, and check HEPA filters. Antibiotics and antimycotics at high concentrations can be toxic to some cell lines; therefore, perform a dose-response test to determine the level at which an antibiotic or antimycotic becomes toxic. This is particularly important when using an antimycotic such as Fungizone Reagent or an antibiotic such as tylosin. The following is a suggested procedure for determining toxicity levels and decontaminating cultures: 1. Dissociate, count, and dilute the cells in antibiotic-free medium. Dilute the cells to the concentration used for regular cell passage. 2. Dispense the cell suspension into a multiwell culture plate or several small flasks. Add the antibiotic of choice to each well or flask in a range of concentrations. For example, for Fungizone Reagent, 0.25, 0.50, 1.0, 2.0, 4.0, and 8.0 g/ml. 3. Observe the cells daily for signs of toxicity such as sloughing, appearance of vacuoles, decrease in confluency, and rounding. 4. When the toxic antibiotic level has been determined, culture the cells for 2 to 3 passages using the antibiotic at a concentration 1 to 2 times lower than the toxic concentration. 5. Culture the cells for 1 passage in antibiotic-free medium. 6. Repeat Step 4. 7. Culture the cells in antibiotic-free medium for 4 to 6 passages to determine if the contamination has been eliminated.
Fungizone is a trademark of E.R. Squibb & Sons, LLC.
Appendix I
Use of Antibiotics and Antimycotics*

Antibiotic Fungizone Reagent (Amphotericin B) Gentamicin sulfate Kanamycin sulfate Neomycin sulfate Penicillin G Polymixin B sulfate Streptomycin sulfate Recommended concentration 0.252.5 g/ml 550 g/ml 100 g/ml 50 g/ml 50100 units/ml 100 units/ml 50100 g/ml Antibiotic spectrum Fungi and yeasts Gram-positive and gram-negative bacteria and mycoplasmas Gram-positive and gram-negative bacteria and mycoplasmas Gram-positive and gram-negative bacteria Gram-positive bacteria Gram-negative bacteria Gram-positive and gram-negative bacteria Stability in tissue culture media at 37C 3 days 5 days 5 days 5 days 3 days 5 days 3 days
The decision to use antibiotics to prevent contamination should be based on the individual researchers needs and experience. This table is a general guide for use of gibco antibiotics in tissue culture media. The concentrations given are for tissue culture media containing serum; serum-free media generally require lower concentrations (15 to 110). Also available are solutions that use one or more antibiotics in conjunction with an antimycotic. For all medium types, optimal concentrations of anti biotics and antimycotics should be determined empirically. Fungizone is a trademark of E.R. Squibb & Sons, LLC.
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Conversion Factors, Units, and Abbreviations

Conversion Factors
base mass
1 kb ds DNA (Na+) = 6.6 105 Da 1 kb ss DNA (Na+) = 3.3 105 Da 1 kb ss RNA (Na+) = 3.4 105 Da 1 MDa ds DNA (Na+) = 1.52 kb average mass of dNMP = 330 Da average mass of dNMP bp = 660 Da
Calculations
molarity of a solution = mass of solute (g) [molecular mass of solute (g/mol)] (L of solution) molecular mass of a ds DNA fragment = (number of bp) (660 Da/bp) moles of ends of a ds DNA fragment = 2 (g of DNA)/[molecular mass of DNA (Da)] or = 2 (g of DNA)/[(number of bp) (660 Da/bp)] moles of ends generated by a restriction endonuclease circular DNA: 2 (moles of DNA) (number of sites) linear DNA: 2 (moles of DNA) (number of sites) + 2 (original ends)
base
protein
1 kb coding capacity = 333 amino acids = 40,000 Da protein 10,000 Da protein = ~270 bp DNA 50,000 Da protein = ~1.35 kb DNA 100,000 Da protein = ~2.7 kb DNA average mass of amino acid = 120 Da
Units of radioactivity
1 Ci = 2.2 106 disintegrations per minute = 3.7 104 becquerels 1 becquerel = 1 disintegration per second
mass of nucleic acid
moles of nucleic acid
1 g/ml of nucleic acid = 3.0 M phosphate 1 g of a 1 kb DNA fragment = 1.5 pmol; 3.0 pmol ends 1 pmol of a 1 kb DNA = 0.66 g 1 g pBR322 = 0.36 pmol DNA 1 pmol pBR322 = 2.8 g 35 nmol of a 24-mer = 277 g
Half-life of common radioisotopes

Radioisotope Carbon-14 (14C) Iodine-125 (125I) Phosphorus-32 (32P) Sulfur-35 (35S) Tritium (3H) Half-life 5,730 years 60 days 14.3 days 87.4 days 12.4 years
absorbance of nucleic acid

Nucleic acid
ds DNA ss DNA ss RNA
concentration of nucleic acid

Concentration (g/ml) for 1 A260 unit
50 33 40
Metric prefixes
M = mega = 106 k = kilo = 103 m = milli = 103 = micro = 106 n = nano = 109 p = pico = 1012 f = femto = 1015 a = atto = 1018
mass of protein
10,000 50,000 100,000 150,000
moles of protein
1 g
100 pmol or 6 10 molecules 20 pmol or 1.2 1013 molecules 10 pmol or 6 10 molecules 6.7 pmol or 4 10 molecules
12 12 13
Molecular weight (Da)
1 nmol
10 g 50 g 100 g 150 g
Abbreviations
ds . . . . . . double-stranded (as in ds DNA) ss . . . . . . . single-stranded (as in ss DNA) bp . . . . . . base pair kb . . . . . . kilobase: 1,000 bases or base pairs, as appropriate Da . . . . . . dalton, the unit of molecular mass mol . . . . . mole M . . . . . . . molarity, moles of solute per liter of solution
concentration of protein
Protein IgG IgM IgA protein A avidin streptavidin bovine serum albumin
absorbance of protein
A280 units for 1 mg/ml 1.35 1.2 1.3 0.17 1.5 3.4 0.7
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Genetic Code and Amino Acid Abbreviations

Second position
U
UUU UUC
Appendix I
UUA UUG
} }
Phenylalanine (Phe, F) Leucine (Leu, L)
UCU UCC UCA UCG
} } } }
C
UAU UAC UAA UAG
G
UGU UGC UGA UGG
Serine (Ser, S)
} } } } } } } }
Tyrosine (Tyr, Y) STOP
Cysteine (Cys, C) STOP Tryptophan (Trp, W)
First position
CUU CUC CUA CUG
}
}
Leucine (Leu, L)
CCU CCC CCA CCG
Proline (Pro, P)
CAU CAC CAA CAG AAU AAC AAA AAG
Histidine (His, H) Glutamine (Gln, Q) Asparagine (Asn, N) Lysine (Lys, K)
CGU CGC CGA CGG
}
} }
Arginine (Arg, R)
AUU AUC AUA AUG
Isoleucine (Ile, I)
ACU ACC ACA ACG
Threonine (Thr, T)
AGU AGC AGA AGG
Serine (Ser, S) Arginine (Arg, R)
GUU GUC GUA GUG
Methionine (Met, M) GCU GCC GCA GCG
Valine (Val, V)
Alanine (Ala, A)
GAU GAC GAA GAG
Aspartic acid (Asp, D) Glutamic acid (Glu, E)
GGU GGC GGA GGG
Glycine (Gly, G)
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Notes:
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Appendix II
Terms and Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236 Trademark Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 238
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Appendix II Trademark Information
Terms and Conditions

The following is an abbreviated description of our General Terms and Conditions. The General Terms and Conditions are included with or on Invitrogen invoices or pack slips and control the sale of products by Invitrogen. In the event of any inconsistency between this catalogue and the General Terms and Conditions, then the General Terms and Conditions shall control. Any customer who does not have a copy of our General Terms and Conditions may obtain a copy by contacting Customer Service. Our General Terms and Conditions are also available on our website at www.invitrogen.com. In addition, customers outside the U.S. should contact their local distributor for a copy of the General Terms and Conditions applicable for their local jurisdiction.
Damaged or Incomplete Shipment

If your shipment is visibly damaged, accept the shipment and retain all shipping documents, cartons, and packing materials for inspection. Note on the waybill any visible damage at the time of receipt and report the damage to a Customer Service Representative as soon as possible. You should also call if you find concealed damage or the shipment is incomplete.
Returns
If a Life Technologies tm catalogue product does not conform to its description in this catalogue or specifications in the product insert sheet, let us know. We will credit your account or exchange the product at our option. To return any catalogue product, you must first obtain a return goods authorization (RGA) number. Please contact your Customer Service Representative for instructions. A restocking fee of twenty-five percent (25%) of the amount invoiced will be charged on the return of any product for any reason other than under our warranty described in this catalogue.
Quick Delivery
If you require special delivery, please request this when placing your order. We will make every attempt to get your order to you when you need it. As appropriate, we will arrange and bill for this service accordingly.
Changes or Cancellations
If your catalogue order has not been shipped, you may initiate a request over the phone to change the order or cancel it. Youll need to confirm your request in writing, as well, and may do so by using the appropriate toll-free ordering fax lines.
Our Warranty
Life Technologies tm sells products in good faith, believing that the products we deliver to you, the original purchaser of our products, will match the description you read in our catalogue and the specifications in the product insert sheet. Below is an abbreviated description of our warranty. The complete warranty statement is located on the reverse of Life Technologiestm invoices or packing lists. In summary, if the product is defective or nonconforming, we will either replace the product or refund the purchase price of the product, provided that: (i) if the problem results from incomplete delivery, you advise us within three (3) days of receipt; (ii) you did not cause the problem by altering or misusing the product, or the like; (iii) you are the original purchaser of the product; and you advised Customer Service by calling the appropriate telephone number within the specified time limits, and complied with the instructions received from the Customer Service Representative. OUR WARRANTY IS LIMITED, AND UNDER NO CIRCUMSTANCES WILL WE BE LIABLE FOR ANY INDIRECT, SPECIAL, INCIDENTAL, CONSEQUENTIAL, OR EXEMPLARY DAMAGES IN EXCESS OF THE PURCHASE PRICE PAID. All terms and conditions, including price, are subject to change without notice, unless otherwise specified in our Quotation.
Shipping
Unless we agree otherwise, freight terms are FOB Life Technologiestm shipping point with freight charges added to your invoice as a separate item. We do not require written confirmation of orders, except for standing, blanket, and change orders. For these exceptions, please clearly mark Confirming Do Not Duplicate on your written confirmation. We take great care to choose the most appropriate shipping methods for each catalogue order based on feedback from our customers. We ensure the accuracy of each order, and we schedule each shipment to arrive at your destination during normal business hours. We reserve the right to make delivery in installments, all such installments to be separately invoiced and paid for when due per invoice. If your complete order does not arrive promptly and in optimal condition, please notify a Customer Service Representative.
Special Shipping Conditions or Carriers

We use air freight and surface carriers to ensure prompt and cost-effective delivery of your order. If you require other arrangements, just tell us when you place your order. We ship at room temperature or on dry ice or wet ice, as detailed in the product descriptions in this catalogue.
Acceptance of Products
Unless we receive written notification from you within three (3) days after receipt, your acceptance of the products shipped by us constitutes your assent to the general terms and conditions under which we offer these materials for sale.
236
Patent Warranty and Disclaimer

We warrant to you, our customers, that the manufacture and sale of the products manufactured by or for us without reliance upon instructions, specifications, or other directions provided by you to our knowledge will not infringe the claims of any patent, trademark or copyright of any third party. We do not warrant that the manufacture and sale by us of products manufactured in reliance upon instructions, specifications, or other directions provided by you or your use or resale of the products will not infringe the claims of any patent, trademark, or copyright of any third party.
Note Hazards
Please read the label for information about product hazards. All Life Technologiestm products should be handled only by qualified persons trained in laboratory safety procedures. Some products may present flammable, corrosive, acute and chronic, toxic, reactive, or other hazards. Product-specific hazard warnings are included in this catalog, in product literature, on product labels, and in MSDSs where applicable. All possible hazards of many products may not be known at this time: the absence of a warning must not be interpreted as an indication that the product is safe. Please report any information of new effects (not already described in literature, on labels, or on MSDSs) to Invitrogen Technical Support. Always use good laboratory procedures when handling any products.
Authorized Use of Products

Except as otherwise agreed in writing by an authorized representative of Invitrogen, the purchase of products only conveys to you the non-transferable right to use the products purchased in compliance with the applicable intended use statement, limited use statement, or limited use label license, if any, in our catalogues or on the label or other documentation accompanying the products. Unless otherwise expressly indicated in our catalogues or on the label or other documentation accompanying the products, the products are intended for research use only and are not to be used for any other purposes including, but not limited to, unauthorized commercial purposes, in vitro diagnostic purposes, ex vivo or in vivo therapeutic purposes, investigational use, in foods, drugs or cosmetics of any kind, or for consumption by or use in connection with or administration or application to humans or animals. You acknowledge that the products have not been tested by or for us for safety or efficacy, unless expressly stated in our catalogues or on the label or other documentation accompanying the products. Without limiting the foregoing restrictions, you warrant to us that should you use or sell the products for any use other than research, you shall conduct all necessary tests, comply with all applicable regulatory requirements, issue all appropriate warnings and information to subsequent purchasers and/or users and be responsible for obtaining any required Intellectual Property rights. Please contact the Companys Licensing Development department through any of the contacts listed on the back of this catalogue to obtain information regarding use of products or their components for purposes other than as authorized above.
Appendix II
Product Labeling and Usage

The products listed in the catalogue are for research use only, unless otherwise stated. Products labeled For in vitro diagnostic use are medical devices and as such are required to comply with the Food and Drug Administration Good Manufacturing Practices known as The Quality System Regulation (Code of Federal Regulations Title 21, Part 820). These products may also be used in laboratory research applications. These products are not qualified nor intended for any animal or human therapeutic uses. Inquiries into the use of Life Technologiestm products are welcome and should be directed to Life Technologiestm Technical Support. Requests will be considered on a case-by-case basis as appropriate under applicable government regulations.
By placing an order, you agree to the following:

The purchaser is responsible for compliance with Food and Drug Administration regulations, all state laws, and any other federal or local laws or regulations governing the use of these materials. Invitrogen recommends that the buyer or other user adhere to NIH guidelines published in the Federal Register, Volume 41, No. 131, July 7, 1976, and any amendments thereto. Invitrogen disclaims any and all responsibility for any injury or damage which may occur from the use of these materials, except as provided in the express warranty set forth above.
Please Read the Label

Please note the expiration dates, if any, and follow the proper usage and storage instructions provided on the label or product profile for each product. We cannot be liable for a product after its expiration date or for one that has been misused or has become unusable due to improper storage, handling, or label alterations. In certain circumstances, the label, product profile, or Material Safety Data Sheet (MSDS) contain important information, instructions, or warnings that were not included in this catalogue. Therefore, it is important to review these items carefully.
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Appendix II Trademark Information
Trademark Information
Life Technologiestm has achieved recognition of its trademarks worldwide. Please respect our trademarks; we will vigorously protect their proper usage. All names containing the designation are registered with the U.S. Patent and Trademark Office. Such trademarks may or may not be registered in other countries.
Trademarks of Life Technologiestm and its affiliated companies

293fectin AIM V AlbuMAX AlgiMatrix AmnioMAX Advanced Granulation Technology (AGT) Bac-N-Blue Bac-to-Bac BaculoDirect BLOCK-iT Cellfectin EpiLife Express Five E-Z Hold EZPassage FoamAway FreeStyle Gateway Geltrex Geneticin gibco GlutaMAX High Five iMATCH InsectSelect Invitrogen KaryoMAX KnocKout Lipofectamine MarrowMAX MediaExpressSM MesenPRO MesenPRO RS Neurobasal Oligofectamine One Shot One Shot OptiCHO OptiMab Opti-MEM OptiPRO pAd/BLOCK-iT pAd/PL-DEST PB-MAX PD-Direct pcDNA PLUS pOptiVEC Recovery Revolution rProtease SimCell Sf-900 Stak Pack StemPro SynerGy Synth-a-Freeze TA Cloning Tech-Line Tech OnLine TOPO TrypLE ViraPower Voyager
Other companies trademarks

C-flex (Consolidated Polymer Technologies) Colcemid (Ciba Ltd.) Cellbag (Wave Biotech) Corning (Corning Inc.) Fungizone (E.R. Squibb & Sons, L.L.C.) Matrigel (Becton Dickinson) PER.C6 (Crucell Holland B.V.) Pluronic (BASF Corporation) Solvay (Solvay SA) Tween (ICI Americas, Inc.) ViCell Cell Counter (Beckman Coulter) Zeocin (CAYLA) Lab Armor
238
Index
Alphabetical Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 240 Catalogue Number Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 246
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Alphabetical Index
Page 250X Cholesterol Lipid Concentrate . . . . . .106, 163, 166 293 Expression Products . . . . . . . . . . . . . . . .93 293 Cells . . . . . . . . . . . . . . . . . . . . . 177 293 SFM II . . . . . . . . . . . . . . . . . . . . . .93 CD 293 AGT . . . . . . . . . . . . . . . . . . . .93 FreeStyle 293 Expression Medium . . . . . . . . . .93 FreeStyle 293 Expression System . . . . . . . . . .94 293fectin Transfection Reagent . . . . . . . . . . . 133 2-Mercaptoethanol (1,000X), liquid . . . . . . . . . . 168 3D Matrices and Scaffolds . . . . . . . . . . . . . . 88 AlgiMatrix 3D Culture System . . . . . . . . . . . . 6 4% Agarose Gel . . . . . . . . . . . . . . . . . . . 103 Page Penicillin-Streptomycin . . . . . . . . . . . . . . 154 Penicillin-Streptomycin-Glutamine . . . . . . . . . 154 Penicillin-Streptomycin-Neomycin (PSN) . . . . . . 155 Polymyxin B Sulfate . . . . . . . . . . . . . . . . 155 Streptomycin Sulfate. . . . . . . . . . . . . . . . 155 Zeocin . . . . . . . . . . . . . . . . . . . . . 155 Antifoaming and Defoaming Reagents . . . . . . . . 156 FoamAway Irradiated . . . . . . . . . . . . . . . 156 FoamAway Irradiated AOF . . . . . . . . . . . . 156 Attachment Factors . . . . . . . . . . . . . . . . . 162 Fibronectin, Human Plasma . . . . . . . . . . . . 162 Natural Mouse Laminin . . . . . . . . . . . . . . 162 Page Collagenase, lyophilized . . . . . . . . . . . . . . 158 Collagenase Type I . . . . . . . . . . . . . . . . . 158 Collagenase Type II . . . . . . . . . . . . . . . . 158 Collagenase Type IV . . . . . . . . . . . . . . . . 158 Dispase . . . . . . . . . . . . . . . . . . . . . . 158 TrypLE Select Stable Trypsin Replacement Enzyme (1X), liquid . . . . . . . . . 159 TrypLE Express, Stable Trypsin Replacement Enzyme (1X), liquid . . . . . . . . . 160 Trypsin, 0.25% (1X), liquid . . . . . . . . . . . . . 160 Trypsin, 2.5% (10X), liquid . . . . . . . . . . . . . 160 Trypsin-EDTA (0.05% Trypsin, EDTA4Na) (1X), liquid . 160 Trypsin-EDTA (0.5% Trypsin, EDTA4Na) (10 X), liquid . 160 Trypsin-EDTA (0.25% Trypsin, EDTA4Na) (1X), liquid . 160 Trypsin (1:250), powder . . . . . . . . . . . . . . 161 Trypsin Inhibitor, soybean . . . . . . . . . . . . . 161 Versene 1:5,000 (1X), liquid Cellfectin Reagent . . . . . . . . . . . . . . . . . 103 Cellfectin II Transfection Reagent . . . . . . . . . . . 133 Cell Line Development Kits . . . . . . . . . . . . . . 180 OptiCHO Express Kit . . . . . . . . . . . . . . . 180 OptiCHO Antibody Express Kit . . . . . . . . . . 181 Cell Therapy Systems (CTS) . . . . . . . . . . . . . . 2 CELLstart Cell Attachment/Expansion Substrate . . . . 4 Cellular Studies . . . . . . . . . . . . . . . . . . . 109 Bronchial Epithelial Products . . . . . . . . . . . . 109 Dermatology Products . . . . . . . . . . . . . . . 110 Hematology-Oncology Products . . . . . . . . . . 110 Hematopoiesis Products . . . . . . . . . . . . . . 113 Hepatocyte Products . . . . . . . . . . . . . . . 113 Human Mammary Epithelial Products . . . . . . . . 114 Neurobiology Products . . . . . . . . . . . . . . 114 Chicken Serum . . . . . . . . . . . . . . . . . . . 192 CHO-S Cells . . . . . . . . . . . . . . . . . . . . . 178 CMRL Medium . . . . . . . . . . . . . . . . . . . .29 CMRL Medium, 1066 (1X) + (10), liquid . . . . . . . .29 CO2-Independent Medium . . . . . . . . . . . . . . .29 CO2-Independent Medium (1X), liquid . . . . . . . .29 Collagenase, lyophilized (Clostridiopeptidase A) . . . . 130 Collagen 1 . . . . . . . . . . . . . . . . . . . . . . 5 Conversion Factors, Units, and Abbreviations. . . . . . 232 Cryopreservation of Mammalian Cells . . . . . . . . . 227 Thawing of Cryopreserved Cells . . . . . . . . . . 227 Cryopreservation Medium . . . . . . . . . . . . . . .29 CTS cytokines and growth factors . . . . . . . . . . . 3 Cytogenetics Products . . . . . . . . . . . . . . . . 126 MarrowMAX . . . . . . . . . . . . . . . . . . . 126 MarrowMAX Bone Marrow Medium . . . . . . . . 126 AmnioMAX . . . . . . . . . . . . . . . . . . . 127 AmnioMAX-II Complete Medium . . . . . . . . . 127 AmnioMAX-C100 Products . . . . . . . . . . . . 127 KaryoMAX . . . . . . . . . . . . . . . . . . . . 127
A
ACK Lysing Buffer . . . . . . . . . . . . . . . . . . 149 Activin A . . . . . . . . . . . . . . . . . . . . . . .14 Actinomycin D . . . . . . . . . . . . . . . . . . . 150 Adaptation of Cell Cultures to a Serum-free Medium . . 224 Adaptation of Cell Cultures to an Alternative Serum . . 225 Adaptation of Cells from Monolayer to Suspension Culture . . . . . . . . . . . . . . . . 225 Advanced DMEM . . . . . . . . . . . . . . . . . . .30 Advanced DMEM/F-12 . . . . . . . . . . . . . . . . .33 Advanced MEM (Minimum Essential Medium), with L-gluatmine . . . . . . . . . . . . . . . . . .40 Advanced RPMI Medium 1640 (1X), liquid . . . . . . . .45 Adenovirus Expression Medium (AEM). . . . . . . . . .96 AIM V Medium CTS . . . . . . . . . . . . . . . . . 3 AIM V Medium (research grade) . . . . . . . 112, 117 AIM V Medium (therapeutic grade) . . . . . . 112, 118 AlbuMAX . . . . . . . . . . . . . . . . . . . . . . 163 AlgiMatrix 3D Culture System . . . . . . . . . . . . . 6 Amino Acids . . . . . . . . . . . . . . . . . . . . 149 GlutaMAX-I Supplement . . . . . . . . . . . . . 149 L-Glutamine . . . . . . . . . . . . . . . . . . . . 149 L-Glutamine-200 mM . . . . . . . . . . . . . . . 149 MEM Amino Acids Solution. . . . . . . . . . . . . 149 AmnioMAX . . . . . . . . . . . . . . . . . . . . 127 Antibiotic-Antimycotic (100X), liquid . . . . . . . . . 129 Antibiotics and Antimycotics . . . . . . . . . . . . . 150 Actinomycin D . . . . . . . . . . . . . . . . . . 150 Ampicillin . . . . . . . . . . . . . . . . . . . . . 150 Antibiotic-Antimycotic . . . . . . . . . . . . . 129, 150 Blasticidin S HCl . . . . . . . . . . . . . . . . . . 151 Puromycin . . . . . . . . . . . . . . . . . . . . 151 Carbenicillin. . . . . . . . . . . . . . . . . . . . 151 Geneticin Selective Antibiotic . . . . . . . . . . . 152 Gentamicin Reagent . . . . . . . . . . . . . . . . 153 Hygromycin B . . . . . . . . . . . . . . . . . . . 153 Kanamycin Sulfate . . . . . . . . . . . . . . . . . 153 Mycophenolic Acid . . . . . . . . . . . . . . . . 154 Neomycin Sulfate . . . . . . . . . . . . . . . . . 154
B
Bags . . . . . . . . . . . . . . . . . . . . . . . . 206 Gibco Universal Bags . . . . . . . . . . . . . . . 206 Prefilled Wave bag . . . . . . . . . . . . . . . . . 207 Balanced Salt Solutions . . . . . . . . . . . . . . . 138 Balanced Salt Solutions Preparation . . . . . . . . . . 142 Balanced Salt Solutions Formulations . . . . . . . . . 143 Basal Medium Eagle (BME) . . . . . . . . . . . . . . .28 BGJb Medium (Fitton-Jackson Modification) . . . . . . .28 Biological Buffers . . . . . . . . . . . . . . . . . . 157 HEPES Buffer Solution (1 M) . . . . . . . . . . . . 157 HEPES Ultrapure . . . . . . . . . . . . . . . . . . 157 Sodium Bicarbonate Solution, 7.5% . . . . . . . . . 157 Blasticidin S HCI . . . . . . . . . . . . . . . . . 123, 151 BMP-4 . . . . . . . . . . . . . . . . . . . . . . . .14 BMP-7 . . . . . . . . . . . . . . . . . . . . . . . .14 Bovine and Rat Tail Collagen . . . . . . . . . . . . . . 5 Bovine Sera . . . . . . . . . . . . . . . . . . . . . 191 Newborn Calf Sera . . . . . . . . . . . . . . . . . 191 Bovine Serum, heat-inactivated . . . . . . . . . . . 191 Donor Bovine Serum. . . . . . . . . . . . . . . . 191 Donor Bovine Serum, with iron . . . . . . . . . . . 191 Brinsters BMOC-3 Medium . . . . . . . . . . . . . . .29 Bronchial Epithelial Products . . . . . . . . . . . . . 109 Buffer Systems for Cell Culture . . . . . . . . . . . . 219
C
Carbenicillin, Disodium Salt . . . . . . . . . . . . . . 151 CD DG44 Medium . . . . . . . . . . . . . . . . . . 180 CD Hybridoma Medium (1X), liquid . . . . . . . . . . 106 CD Hybridoma AGT. . . . . . . . . . . . . . . . . 106 CD OptiCHO Medium . . . . . . . . . . . . . . . 180 Cell Culture basics . . . . . . . . . . . . . . . . . . 210 Cell Culture Products Overview . . . . . . . . . . . . . ii Cell Culture Reagents . . . . . . . . . . . . . . . . 148 Other Cell Culture Reagents . . . . . . . . . . . . 168 Cell Culture Reagent Formulations . . . . . . . . . 169 Cell Dissociation Products . . . . . . . . . . . . . . 158 Cell Dissociation Buffers . . . . . . . . . . . . . . 158
240
Page PB-MAX Karyotyping Medium . . . . . . . . . . . 127 KaryoMAX Colcemid Solution, liquid (10 g/ml), in HBSS . . . . . . . . . . . . 127 Cytogenetics: Related Products . . . . . . . . . . . . 128 Gurr Buffer Tablets (pH 6.8) . . . . . . . . . . . . . 128 Phytohemagglutinin (M Form) (PHA), liquid . . . . . 128 Potassium Chloride Solution, 0.075 M . . . . . . . . 128
Page Insect Cell Expression Products . . . . . . . . . . . .98 Customized Insect Medium Products . . . . . . . . 101 Insect Medium Supplements . . . . . . . . . . . . 103 Hybridoma and Monoclonal Antibody Products . . . 106 Customized Hybridoma Medium Products . . . . . 107 E-Z Hold Plastic Bottles . . . . . . . . . . . . . . . .49
Page Geltrex Reduced Growth Factor Basement Membrane Matrix . . . . . . . . . . . . . . . . . . . . . . 5 Bovine and Rat Tail Collagen I . . . . . . . . . . . . . 5 Gibco CTS Immunotherapy products . . . . . . . . . 3 AIM V Medium CTSTherapeutic-grade, serum-free cell expansion medium . . . . . . . . . 3 OpTmizer CTS T-Cell Expansion SFM . . . . . . . . 3 CTS cytokines and growth factors . . . . . . . . . . 3 DPBS CTS . . . . . . . . . . . . . . . . . . . . . 3 Dynabeads CD3/CD28 CTS . . . . . . . . . . . . 3 DynaMag CTS . . . . . . . . . . . . . . . . . . 3 Interleukin 4 CTS . . . . . . . . . . . . . . . . . . 3 Gibco CTS products for stem cells . . . . . . . . . . 4 Gibco LIF Recombinant . . . . . . . . . . . . . . . .13 Gibco Recombinant Growth Factors for use in stem cell applications . . . . . . . . . . . . . . . . . . . . .14 Gibco Recombinant human FGF-basic . . . . . . . . .12 Gibco Universal Bags . . . . . . . . . . . . . . . . 204 Prefilled Wave Bag . . . . . . . . . . . . . . . . . 207 Glasgow Minimum Essential Medium . . . . . . . . . .35 Glasgow Minimum Essential Medium . . . . . . . . .35 (G-MEM, BHK-21) (1X), liquid GlutaMAX Media at a glance . . . . . . . . . . . . .26 GlutaMAX-I Supplement . . . . . . . . . .123, 130, 149 Goat Serum . . . . . . . . . . . . . . . . . . . . . 192 Graces Insect Cell Culture Media . . . . . . . . . . . .35 Graces Insect Cell Culture Medium, supplemented (1X), liquid . . . . . . . . . . . . . .35 Graces Insect Cell Culture Medium, supplemented (2X), liquid . . . . . . . . . . . . . .35 Graces Insect Cell Culture Medium, unsupplemented (1X), liquid . . . . . . . . . . . . .35 Graces Insect Cell Culture Medium, powder . . . . . .35 Growth factors, Cytokines and Chemokines with high bioactivity . . . . . . . . . . . . . . . . . . . . .10 Growth Supplements . . . . . . . . . . . . . . . . 163 250X Cholesterol Lipid Concentrate . . . . . . . . . 163 Chemically Defined Lipid Concentrate . . . . . . . 163 AlbuMAX I . . . . . . . . . . . . . . . . . . . . 163 AlbuMAX II . . . . . . . . . . . . . . . . . . . . 163 Bovine Albumin Fraction V Solution (7.5%). . . . . . 163 B-27 Serum-free Supplements . . . . . . . . . . . 164 G-5 Supplement (100X), liquid . . . . . . . . . . . 164 Insulin, Human Recombinant, Zinc . . . . . . . . . 164 Insulin-Transferrin-Selenium Supplements (ITS) . . . 164 Insulin-Transferrin-Selenium-A Supplement (100X) . . 164 Insulin-Transferrin-Selenium-X Supplement (100X) . . 165 Lactalbumin Hydrolysate, powder (extra soluble). . . 165 N-2 Supplement (100X), liquid . . . . . . . . . . . 165 Transferrin, Bovine (APO form), lyophilized . . . . . . 165 Transferrin, Bovine (holo form), lyophilized . . . . . . 165 Transferrin, Bovine (partially iron loaded), lyophilized . 165
D
Decontaminating Cultures with Antibiotics and Antimycotics . . . . . . . . . . . . . . . . . 231 Use of Antibiotics and Antimycotics . . . . . . . . . 231 Defined Keratinocyte-SFM System . . . . . . . . . . 110 Dissociation of Cells from Primary Tissue . . . . . . . . 228 Dissociation of Cells from Culture Vessels . . . . . . . 229 Distilled Water . . . . . . . . . . . . . . . . . . . 161 Dermatology Products . . . . . . . . . . . . . . . . 110 Keratinocyte Starter Kit. . . . . . . . . . . . . . . 110 Keratinocyte-SFM (1X), liquid . . . . . . . . . . . . 110 Keratinocyte-SFM (1X), liquid (without CaCl2) . . . . 110 DG44 Cells . . . . . . . . . . . . . . . . . . . . . 178 D-Glucose (dextrose) . . . . . . . . . . . . . . . . . 168 D.Mel-2 Cells . . . . . . . . . . . . . . . . . . . . 174 DPBS CTS . . . . . . . . . . . . . . . . . . . . . . 3 Dulbeccos Modified Eagle Media (DMEM) . . . . . . . .30 Advanced DMEM (1X), liquid . . . . . . . . . . . . .30 DMEM (1X), liquid (high glucose) . . . . . . . . . . .30 DMEM (1X), liquid (low glucose) . . . . . . . . . . .32 DMEM (1X), liquid (no glucose) . . . . . . . . . . . .32 DMEM, powder (low glucose) . . . . . . . . . . . .32 Advanced DMEM/F-12 (1X), liquid, 1:1 . . . . . . . . .33 DMEM/F-12 (1X), liquid, 1:1 . . . . . . . . . . . . . .33 Dulbeccos Phosphate-Buffered Salines . . . . . . . . 138 Dulbeccos Phosphate-Buffered Saline (DPBS) (1X), liquid . . . . . . . . . . . . . . . . . . . . 138 Dulbeccos Phosphate-Buffered Saline (DPBS) (10X), liquid . . . . . . . . . . . . . . . . . . . . 139 Dulbeccos Phosphate-Buffered Saline (DPBS), powder . . . . . . . . . . . . . . . . . . . . . . 139 Dynabeads CD3/CD28 CTS . . . . . . . . . . . . . 3 DynaMag CTS . . . . . . . . . . . . . . . . . . . 3
F
F-10 Nutrient Mixtures . . . . . . . . . . . . . . . . .34 F-10 Nutrient Mixture (Ham) (1X), liquid . . . . . . . .34 F-12 Nutrient Mixtures . . . . . . . . . . . . . . . . .34 F-12 Nutrient Mixture (Ham) (1X), liquid . . . . . . . .34 F-12 Nutrient Mixture (Ham), powder . . . . . . . . .34 F-12K Nutrient Mixture, Kaighns Modification (1X), liquid . . . . . . . . . . .34 FBS Selection Guide . . . . . . . . . . . . . . . . . 187 FBS Quality Control Test Comparison . . . . . . . . . 193 Other Sera Quality Control Test Comparison . . . . . 194 Fetal Bovine Sera (FBS) and other Sera for Cell Culture . 186 General Animal Sera Information . . . . . . . . . . 186 The iMATCH MPA Sera Lot Matching Tool. . . . . . 186 Fetal Bovine Sera (FBS) . . . . . . . . . . . . . . . . 189 qualified . . . . . . . . . . . . . . . . . . . . . 188 qualified, heat-inactivated . . . . . . . . . . . . . 188 qualified, gamma irradiated . . . . . . . . . . . . 189 dialyzed . . . . . . . . . . . . . . . . . . . . . 189 charcoal stripped . . . . . . . . . . . . . . . . . 189 ES cell qualified . . . . . . . . . . . . . . . . . . 189 MSC qualified . . . . . . . . . . . . . . . . . . . 190 ultra-low IgG FBS . . . . . . . . . . . . . . . . . 190 FGF-basic . . . . . . . . . . . . . . . . . . . . . . .12 Fibronectin, Human Plasma. . . . . . . . . . . . . . 162 Final Product Quality Control . . . . . . . . . . . . 196 FoamAway Irradiated . . . . . . . . . . . . . . . . 156 FoamAway Irradiated AOF . . . . . . . . . . . . . 156 Formulas and Formula Weights for Commonly Used Medium Components . . . . . . . . . . . . . . . 216 FreeStyle MAX Reagent . . . . . . . . . . . . . . . 132 FreeStyle 293 Expression Medium . . . . . . . . . . 177 Fungizone Antimycotic, liquid . . . . . . . . . . . . 129
Alphabetical Index
E
Earles Balanced Salt Solution . . . . . . . . . . . . . 139 Earles Balanced Salt Solution (EBSS) (1X), liquid . . . 139 Expression Products . . . . . . . . . . . . . . . . . 89 CHO Expression Products . . . . . . . . . . . . . .89 Customized CHO Medium Products . . . . . . . . . .90 DG44 Expression Products . . . . . . . . . . . . . .91 293 Expression Products . . . . . . . . . . . . . . .93 Virus Production Medium Products . . . . . . . . . .94 Viral Expression Products . . . . . . . . . . . . . . .97
G
Geltrex Reduced Growth Factor Basement Membrane Matrix . . . . . . . . . . . . . . . . . . . . . . . 5 Genetic Code and Amino Acid Abbreviations . . . . . 233 Geneticin Selective Antibiotic . . . . . . . . . . . . 122 Geneticin Selective Antibiotic, liquid . . . . . . . . . 152 Gentamicin Reagent Solution (10 mg/ml), liquid . . . . 129 Gentamicin Reagent Solution, liquid . . . . . . . . . 153 Geneticin Selective Antibiotic, powder . . . . . . . . 153 Gibco Cell Therapy Systems (CTS) . . . . . . . . . . 2 CELLstart Cell Attachment/Expansion Substrate . . . 4
241
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Alphabetical Index
Page Page Yeastolate Ultrafiltrate (50X), liquid . . . . . . . . . 167 Interleukin 4 CTS . . . . . . . . . . . . . . . . . . . 3 IPL-41 Insect Medium . . . . . . . . . . . . . . . . .36 Iscoves Modified Dulbeccos Media . . . . . . . . . . .36 Iscoves Modified Dulbeccos Medium (IMDM) (1X), liquid . . . . . . . . . . . . . . . . . .36 Iscoves Modified Dulbeccos Medium (IMDM), powder . . . . . . . . . . . . . . . . . . . . . . .36 Advanced Minimum Essential Medium (MEM) (1X), liquid (with L-glutamine) . . . . . . . . . . . . . . .40 Minimum Essential Medium (MEM) with Earles Salts (with L-glutamine) . . . . . . . . . .40 Minimum Essential Medium (MEM) with Earles Salts (without L-glutamine) . . . . . . . .41 Minimum Essential Medium (MEM) with Hanks Salts, liquid (with L-glutamine). . . . . . .42 Modified Eagle Media . . . . . . . . . . . . . . . . .42 Mycophenolic Acid . . . . . . . . . . . . . . . . . 154 Mycoplasma Detection . . . . . . . . . . . . . . . 167 Yeast Extraction Solution . . . . . . . . . . . . . . 167 Page
H
Hanks Balanced Salt Solutions . . . . . . . . . . . . 140 Hanks Balanced Salt Solution (HBSS) (1X), liquid . . . 140 Hanks Balanced Salt Solution (HBSS) (10X), liquid . . 140 HAT Supplement (50X), liquid . . . . . . . . . . . . . 107 Gibco Hepatocytes . . . . . . . . . . . . . . . . . 182 Hematopoiesis Products . . . . . . . . . . . . . . . 113 StemPro-34 SFM (1X), liquid . . . . . . . . . . . . 113 Macrophage-SFM (1X), liquid . . . . . . . . . . . . 113 Hepatocyte Products . . . . . . . . . . . . . . . . . 113 Hepatozyme-SFM (1X), liquid . . . . . . . . . . . . 113 Liver Digest Medium (1X), liquid . . . . . . . . . . 113 Liver Perfusion Medium (1X), liquid . . . . . . . . . 113 Hepatocyte Wash Medium . . . . . . . . . . . . . 113 HEPES Buffer Solution (1 M) . . . . . . . . . . . . . . 157 HEPES Media at a glance . . . . . . . . . . . . . . . .27 HEPES Ultrapure . . . . . . . . . . . . . . . . . . . 157 High Five Cells . . . . . . . . . . . . . . . . . . . 174 Horse Sera . . . . . . . . . . . . . . . . . . . . . 192 Horse Serum . . . . . . . . . . . . . . . . . . . 192 Horse Serum, heat-inactivated . . . . . . . . . . . 192 Human Mammary Epithelial Products . . . . . . . . . 114 HuMEC Medium . . . . . . . . . . . . . . . . . . 114 Human proteins . . . . . . . . . . . . . . . . . . . .15 HT Supplement (100X), liquid . . . . . . . . . . . . . 107 Hybridoma and Monoclonal Antibody Products . . . . 106 Hybridoma Reagents . . . . . . . . . . . . . . . . 166 250X Cholesterol Lipid Concentrate . . . . . . . . . 166 Chemically Defined Lipid Concentrate . . . . . . . 166 HAT Supplement (50X), liquid. . . . . . . . . . . . 166 HT Supplement (100X), liquid. . . . . . . . . . . . 166 Hybridoma-SFM (1X), liquid . . . . . . . . . . . . . . 106 Hygromycin B . . . . . . . . . . . . . . . . . . 123, 153
K
Kanamycin Sulfate . . . . . . . . . . . . . . . . . . 153 Kanamycin Sulfate (100X), liquid . . . . . . . . . . . 153 KaryoMAX . . . . . . . . . . . . . . . . . . . . . 127 KnockOut Serum Replacement . . . . . . . . . . . 120 KnockOut DMEM . . . . . . . . . . . . . . . . . . 121
N
N-2 Supplement . . . . . . . . . . . . . . . . . 115, 117 Natural Mouse Laminin . . . . . . . . . . . . . . . . 6 Neomycin Sulfate . . . . . . . . . . . . . . . . . . 154 Neurobasal Medium . . . . . . . . . . . . . . . . 117 Neurobiology Products . . . . . . . . . . . . . . . . 114 Neurobasal Media . . . . . . . . . . . . . . . . 114 B-27 Serum-free Supplements . . . . . . . . . . . 115 G-5 Supplement (100X), liquid . . . . . . . . . . . 115 N-2 Supplement (100X), liquid . . . . . . . . . . . 115 Brain-Derived Neurotrophic Factor (BDNF), Human, Recombinant . . . . . . . . . . . . . . . . . . 115 Nerve Growth Factor 2.5S (NGF 2.5S), Murine, Natural 116 Nerve Growth Factor 7S (NGF 7S), Murine, Natural . . 116 ViraPower Lentiviral Expression System . . . . . . 116 Newborn Calf Sera . . . . . . . . . . . . . . . . . . 191 Newborn Calf Serum. . . . . . . . . . . . . . . . 191 Newborn Calf Serum, heat-inactivated . . . . . . . 191
L
Lab Armor Beads . . . . . . . . . . . . . . . . . . 208 Lamb Serum . . . . . . . . . . . . . . . . . . . . 192 Leibovitzs L-15 Media . . . . . . . . . . . . . . . . .37 L-Glutamine . . . . . . . . . . . . . . . . . . . . . 149 L-Glutamine-200 mM, liquid . . . . . . . . . . . . . 149 LHC Medium (1X), liquid . . . . . . . . . . . . . . . 109 LIF Recombinant . . . . . . . . . . . . . . . . . . .13 Lipofectamine Transfection Reagent . . . . . . . . . 133 Lipofectamine 2000 Transfection Reagent . . . . 132, 133 Lipofectamine 2000 CD Transfection Reagent . . . . 132 Lipofectamine LTX Transfection Reagent . . . . . . . 132
M
Mammalian Cells . . . . . . . . . . . . . . . . . . 177 293 Cells . . . . . . . . . . . . . . . . . . . . . 177 FreeStyle 293 Expression Medium . . . . . . . . . 177 CHO-S Cells . . . . . . . . . . . . . . . . . . . . 178 DG44 Cells . . . . . . . . . . . . . . . . . . . . 178 Primary Human Keratinocytes . . . . . . . . . . . 179 MarrowMAX . . . . . . . . . . . . . . . . . . . . 126 McCoys 5A Medium (Modified) . . . . . . . . . . . . .37 MCDB 131 Medium . . . . . . . . . . . . . . . . . .37 Media 199 . . . . . . . . . . . . . . . . . . . . . . .38 Medium 199 with Earles Salts . . . . . . . . . . . .38 Medium 199 (10X), liquid. . . . . . . . . . . . . . .38 Medium 199, powder . . . . . . . . . . . . . . . .38 Medium 199 with Hanks Salts . . . . . . . . . . . .38 Media Formulations . . . . . . . . . . . . . . . . . .50 Medium NCTC-109 . . . . . . . . . . . . . . . 42, 107 MEM Amino Acids Solution (50X), liquid . . . . . . . . 149 MEM Non-essential Amino Acids Solution 10 mM (100X), liquid . . . . . . . . . . . . . . . . . . . . . . . 149 MEM Vitamin Solution (100X), liquid . . . . . . . . . . 168 MesenPRO RS Medium . . . . . . . . . . . . . . . 118 Minimum Essential Media (MEM) . . . . . . . . . . . .39
O
OptiCHO Express Kit . . . . . . . . . . . . . . . . 180 OptiCHO Antibody Express Kit . . . . . . . . . . . . 181 Opti-MEM I Reduced-Serum Media . . . . . . . . . . .43 Opti-MEM I Reduced-Serum Medium (1X), liquid . . .43 Opti-MEM I Reduced-Serum Medium (1X), liquid (without phenol red). . . . . . . . . . . . . . . . .43 Opti-MEM I Reduced-Serum Medium (1X), liquid (with GlutaMAX-I) . . . . . . . . . . . . . . . . .43 Opti-MEM I Reduced-Serum Medium, powder . . . .43 Lipofectamine 2000 Transfection Reagent . . . . . .43 OptiPRO SFM . . . . . . . . . . . . . . . . . . . 180 OpTmizer CTS T-Cell Expansion SFM . . . . . . . . . 3 Ordering and Technical Support . . . . . . . . . . . . v
I
IL-17F . . . . . . . . . . . . . . . . . . . . . . . .15 iMATCH MPA Sera Lot Matching Tool . . . . . . . . 186 Improved MEM Zn++ Option (Richters Modification) . . .36 Insect Cells . . . . . . . . . . . . . . . . . . . . . 174 D.Mel-2 Cells . . . . . . . . . . . . . . . . . . . 174 High Five Cells . . . . . . . . . . . . . . . . . . 174 Sf9 Cells . . . . . . . . . . . . . . . . . . . . . 175 Sf21 Cells . . . . . . . . . . . . . . . . . . . . . 175 Insect Cell Expression Products . . . . . . . . . . . . .98 Sf-900 III SFM . . . . . . . . . . . . . . . . . . .99 Sf-900 II SFM . . . . . . . . . . . . . . . . . . . .99 Insect Medium Supplements . . . . . . . . . . . 103, 167 4% Agarose Gel . . . . . . . . . . . . . . . . . . 167 Chemically Defined Lipid Concentrate . . . . . . . 167 Lactalbumin Hydrolysate, powder (extra soluble). . . 167 Pluronic F-68, 10% (100X) . . . . . . . . . . . . . 167
P
pcDNA3.3-TOPO TA Cloning Kit . . . . . . . . . . 181 PD-Direct Bioprocess Services . . . . . . . . . . . . 209 Penicillin-Streptomycin, liquid . . . . . . . . . . 129, 154 Penicillin-Streptomycin-Glutamine (100X), liquid . . . . 154
242
Page Penicillin-Streptomycin-Neomycin (PSN) Antibiotic Mixture (100X), liquid . . . . . . . . . . 155 Performance Tests: Fetal Bovine Sera (FBS) . . . . . . . 197 PFHM-II Protein-free Hybridoma Medium (1X), liquid . . 107 Phenol Redfree Media at a glance . . . . . . . . . . .28 Phosphate-Buffered Saline (PBS) . . . . . . . . . . . 141 Phosphate-Buffered Saline (PBS) 7.2 (1X), liquid . . . 141 Phosphate-Buffered Saline (PBS) 7.2 (10X), liquid . . . 141 Phosphate-Buffered Saline (PBS) 7.4 (1X), liquid . . . 141 Phosphate-Buffered Saline (PBS) 7.4 (10X), liquid . . . 141 Phytohemagglutinin (M Form) (PHA), liquid . . . . . . 168 Pluronic F-68, 10% (100X) . . . . . . . . . . . . . . 103 PLUS Reagent . . . . . . . . . . . . . . . . . . . 133 Polymyxin B Sulfate . . . . . . . . . . . . . . . . . 155 Porcine Serum . . . . . . . . . . . . . . . . . . . . 192 pOptiVEC-TOPO vector. . . . . . . . . . . . . . . 180 Prefilled Wave Cellbag . . . . . . . . . . . . . . . . 207 Presto Blue . . . . . . . . . . . . . . . . . . . . 170 Primary Human Keratinocytes . . . . . . . . . . . . 179 Protein A Agarose . . . . . . . . . . . . . . . . . . 108 Publication References . . . . . . . . . . . . . . . . .20 Puromycin . . . . . . . . . . . . . . . . . . . . . 151
Page Sf-900 II SFM . . . . . . . . . . . . . . . . . . . . .99 Sodium Bicarbonate Solution, 7.5% (w/v) . . . . . . . 130 Sodium Pyruvate Solution 100 mM (100X), liquid . . . . 168 Special Proccessing of Sera . . . . . . . . . . . . . . 200 Specialty FBS Selection Guide. . . . . . . . . . . . . 189 Specialty Media . . . . . . . . . . . . . . . . . . . .85 Specialty Media and Applications . . . . . . . . . . . 219 Specialty Reagents . . . . . . . . . . . . . . . . . .87 Stability of GlutaMAX-I vs. L-Glutamine . . . . . . . . 226 Stak Pack Accessories . . . . . . . . . . . . . . . .49 Stem Cell Research Products . . . . . . . . . . . . . 117 B-27 Supplement . . . . . . . . . . . . . . . . . 117 N-2 Supplement . . . . . . . . . . . . . . . . . . 117 Neurobasal Medium . . . . . . . . . . . . . . . 117 Neurobasal-A Medium . . . . . . . . . . . . . . 117 AIM V Media (research grade) . . . . . . . . . . . 117 AIM V Media (therapeutic grade) . . . . . . . . . . 118 MesenPRO RS Medium . . . . . . . . . . . . . . 118 StemPro hESC SFM . . . . . . . . . . . . . . . . 119 StemPro-34 SFM . . . . . . . . . . . . . . . . . 120 DMEM/F-12 . . . . . . . . . . . . . . . . . . . . 120 KnockOut Serum Replacement . . . . . . . . . . 120 KnockOut DMEM . . . . . . . . . . . . . . . . . 121 Geneticin Selective Antibiotic . . . . . . . . . . . 122 Blasticidin S HCI . . . . . . . . . . . . . . . . . . 123 Hygromycin B . . . . . . . . . . . . . . . . . . . 123 GlutaMAX-I Supplement . . . . . . . . . . . . . 123 Fetal Bovine Serum, ES Cell-Qualified . . . . . . . . 123 CO2-Independent Medium . . . . . . . . . . . . . 123 Basic Fibroblast Growth Factor (bFGF), Human, Recombinant . . . . . . . . . . . . . . . . . . . 124 Insulin-like Growth Factor-I (IGF-I), Human, Recombinant . . . . . . . . . . . . . . . 124 Platelet Derived Growth Factor BB (PDGF BB), Human, Recombinant . . . . . . . . . . . . . . . 124 Streptavidin Agarose . . . . . . . . . . . . . . . . . 108 Streptomycin Sulfate . . . . . . . . . . . . . . . . . 155 Supply Centre by Life Technologiestm . . . . . . . . . 213 Swine proteins . . . . . . . . . . . . . . . . . . . .19
Page Lipofectamine Transfection Reagent . . . . . . . . 133 Lipofectamine 2000 Transfection Reagent . . . . . 133 Troubleshooting: Cell Culture . . . . . . . . . . . . . 218 Trypan Blue Stain . . . . . . . . . . . . . . . . . . 168
UZ
Ultralow IgG FBS . . . . . . . . . . . . . . . . . . . 190 Vascular-Endothelial Products. . . . . . . . . . . . . 124 Basic Fibroblast Growth Factor (bFGF), Human, Recombinant . . . . . . . . . . . . . . . 125 Fibronectin, Human Plasma . . . . . . . . . . . . 125 Human Endothelial-SFM . . . . . . . . . . . . . . 124 Versene . . . . . . . . . . . . . . . . . . . . . . . 161 Viral Expression Products . . . . . . . . . . . . . . . .97 ViraPower Adenoviral Expression System . . . . . . .97 ViraPower Lentiviral Expression System . . . . . . 116 Water, distilled . . . . . . . . . . . . . . . . . . . . 130 Waymouths MB 752/1 Medium . . . . . . . . . . . .46 WFI for Cell Culture (Water for Injection) . . . . . . . 161 Williams Medium E . . . . . . . . . . . . . . . . . .46 Yeastolate Ultrafiltrate (50X), liquid . . . . . . . . . . 103 Zeocin . . . . . . . . . . . . . . . . . . . . . . . 155
Alphabetical Index
R
Rabbit Serum . . . . . . . . . . . . . . . . . . . . 192 Rat proteins . . . . . . . . . . . . . . . . . . . . . .18 Recombinant human FGF-basic . . . . . . . . . . . .12 Recombinant Protein G (rProtein G) Agarose . . . . . . 108 Recovery Freezing Medium . . . . . . . . . . . . . .29 Reduced-serum Media at a glance . . . . . . . . . . .25 RPMI Media 1640 . . . . . . . . . . . . . . . . . . .44 RPMI Medium 1640 (1X), liquid . . . . . . . . . . . .45 RPMI Medium 1640, powder . . . . . . . . . . . . .45 RPMI Medium 1640 (1X), liquid (Folate-free) . . . . . 130
S
Schneiders Drosophila Medium . . . . . . . . . . . .46 Serum-Free, Protein-free, and Chemically Defined Media .85 Serum-free Media and Reagents . . . . . . . . . . . .86 Serum-free Media and Applications . . . . . . . . . . 220 Hybridoma Culture . . . . . . . . . . . . . . . . 220 Chinese Hamster Ovary (CHO) Cell Culture . . . . . 221 Human Embryonic Kidney (293) Cell Culture . . . . . 221 Mammalian Cell Culture for Virus Production . . . . 221 Insect Cell Culture . . . . . . . . . . . . . . . . . 222 Neuronal Cell Culture . . . . . . . . . . . . . . . 222 Blood and Bone Marrow Culture . . . . . . . . . . 223 Stem Cell Culture . . . . . . . . . . . . . . . . . 223 Other Mammalian Cell Culture . . . . . . . . . . . 223 Sf9 Cells . . . . . . . . . . . . . . . . . . . . . . . 175 Sf21 Cells . . . . . . . . . . . . . . . . . . . . . . 175 Sf-900 III SFM . . . . . . . . . . . . . . . . . . . . .99
T
Thawing of Cryopreserved Cells. . . . . . . . . . . . 227 Thrombopoietin (TPO) . . . . . . . . . . . . . . . . .14 Transfection Reagents . . . . . . . . . . . . . . . . 132 Lipofectamine 2000 . . . . . . . . . . . . . . . 132 Lipofectamine LTX . . . . . . . . . . . . . . . . 132 FreeStyle MAX Reagent . . . . . . . . . . . . . . 132 293fectin Transfection Reagent . . . . . . . . . . 133 Lipofectamine RNAiMAX Transfection Reagent . . . 133 Cellfectin Transfection Reagent . . . . . . . . . . 133 PLUS Reagent . . . . . . . . . . . . . . . . . . 133
243
www.invitrogen.com
Alphabetical Index
Notes:
244
Notes:
Alphabetical Index 245

www.invitrogen.com
Catalogue Number Index

Cat. no.
402-03D . . 006-0096BA 006-0096BC 0070037BA . 0070049BA . 0070049BB . 087-0112BK . 087-0112BK . 087-0112BK . 087-0112DK 10010-001 . 10010-002 . 10010-015 . 10010-031 . 10010-056 . 10010-072 . 10082-139 . 10082-147 . 10092-013 . 10099-133 . 10099-141 . 10100-139 . 10100-147 . 10106-151 . 10106-169 . 10108-157 . 10108-165 . 10109-155 . 10109-163 . 10131-019 . 10131-027 . 10131-027 . 10131-027 . 10131-035 . 10177-012 . 10265-015 . 10270-098 . 10270-106 . 10362-010 . 10362-100 . 10362-125 . 10370-047 . 10370-070 . 10371-029 . 10372-019 . 10373-017 . 10378-016 . 10439-016 . 10439-024 . 10486-025 . 10500-056 . 10500-064 . 10564-037 . 10564-045 . 10566-032 . 10566-040 . 10569-069 . 10569-077 . 10575-090 . 10576-015 . 10576-015 . 10582-013 . 10639-011 . 10639-011 . 10687-010 . 10687-010 . 10743-001 . 10743-002 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . 3 . . . . 156 . . . . 156 . . 99, 207 . . . . 207 . . 93, 207 . . . . 112 . . . . 118 . . . . . 3 . 3, 112, 118 . . . . 206 . . . . 206 . . . . 141 . . . . 141 . . . . 141 . . . . 141 . . . . 187 . . . . 187 . . . . 128 . . . . 188 . . . . 188 . . . . 188 . . . . 188 . . . . 188 . . . . 188 . . . . 188 . . . . 188 . . . . 189 . . . . 189 . . . . 122 . . . . 122 . . . . 152 . . . . 181 . . . . 152 . . . . 151 . . . . .49 . . . . 188 . . . . 188 . . . . 133 . . . . 103 . . . . 103 . . . . .41 . . . . .41 . . . . 191 . . . . .37 . . . . .36 . . . . 154 . . . . 123 . . . . 123 . . . . 100 . . . . 188 . . . . 188 . . . . 206 . . . . 206 . . . . 206 . . . . 206 . . . . 206 . . . . 206 . . . . 128 . . . . 128 . . . . 168 . . . . 128 . . . . 113 . . . . 120 . . . . 123 . . . . 153 . . . . 206 . . . . 206
Cat. no.
10744-019 10828-028 10829-018 10831-014 10833-010 10833-028 10833-036 10888-022 10888-022 10889-038 10889-038 10889-038 10902-088 10902-096 10902-104 10902-161 10902-179 10902-187 10908-010 10967-032 11003-019 11020-021 11020-039 11021-029 11021-037 11021-045 11039-021 11039-047 11058-021 11058-021 11067-030 11067-030 11105-012 11107-018 11107-047 11108-016 11111-044 11120-037 11126-034 11130-036 11130-077 11140-035 11140-068 11269-016 11279-023 11300-027 11330-099 11330-107 11344-041 11360-039 11360-088 11405-057 11405-081 11415-049 11415-056 11496-015 11497-013 11514-015 11593-027 11595-030 11595-030 11605-045 11605-045 11605-086 11605-086 11619-012 11625-019 11631-017 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . 110 . . 120 . . 121 . . 174 . . .49 . . .49 . . .49 . . 114 . . 117 . . 115 . . 117 . . 164 . . .99 . . .99 . . .99 99, 206 . . 206 99, 206 . . 115 . . 100 . . .49 . . 163 . . 163 . . 163 . . 163 . . 163 . . .33 . . .33 . . .43 . . .95 . . 107 . . 166 . . 165 . . 165 . . 165 . . 165 . . 125 . . 168 . . .29 . . 149 . . 149 . . 149 . . 149 . . 127 . . 106 . . .35 . . 206 33, 206 . . 157 . . 168 . . 168 . . .36 . . 100 . . .37 . . .37 . . 175 . . 176 . . 133 . . 150 . . 100 . . .35 . . 100 . . .35 . . 100 . . .35 . . 178 . . 177 . . 177
Cat. no.
11667-037 11667-037 11668-019 11668-019 11668-019 11668-027 11668-027 11668-027 11668-500 11668-500 11681-020 11686-029 11686-029 11765-047 11800-042 11800-042 11805-017 11811-023 11811-023 11811-031 11811-031 11811-064 11811-064 11811-098 11811-098 11814-019 11815-024 11815-032 11835-063 11860-038 11875-168 11875-176 11879-020 11880-028 11880-028 11900-073 11905-031 11905-031 11905-031 11905-031 11913-019 11913-019 11960-044 11960-085 11965-167 11965-175 11966-025 11971-025 11995-115 11995-123 12000-063 12002-010 12040-077 12045-076 12045-084 12055-083 12055-083 12055-091 12055-091 12065-074 12102 . . 12260-014 12309-019 12309-050 12309-050 12309-050 12332-011 12338-001 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100 .35 132 133 .43 132 133 .43 132 133 .95 .93 .97 .34 165 167 150 122 153 122 153 122 153 122 153 154 153 153 .45 155 206 206 .45 118 .32 .40 103 163 166 167 .93 .96 .30 .30 206 206 .32 .30 206 206 .40 .49 107 106 106 112 117 112 117 113 . 3 126 .94 180 181 .92 179 206
Cat. no.
12338-002 12338-018 12338-018 12338-026 12338-026 12347-019 12347-019 12347-500 12347-750 12348-017 12349-015 12372-017 12372-025 12430-112 12430-120 12440-046 12491-015 12491-023 12492-013 12492-021 12529-012 12529-012 12529-020 12529-020 12531-018 12556-015 12556-023 12557-013 12557-021 12558-011 12559-019 12559-027 12563-011 12563-011 12563-029 12563-029 12566-014 12568-010 12582-011 12585-014 12587-010 12587-010 12587-010 12591-038 12604-013 12604-013 12604-021 12604-021 12604-039 12604-054 12605-010 12605-028 12605-036 12605-093 12610-010 12610-010 12610-010 12618-012 12618-013 12633-012 12633-020 12634-010 12634-028 12648-010 12658-001 12658-019 12658-027 12658-035 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . 206 . . . . . .93 . . . . . .94 . . . . . 177 . . . . . .93 . . . . . 133 . . . . . .94 . . . . . 133 . . . . . 133 . . . . . 114 . . . . . 114 . . . . . 106 . . . . . 106 . . . . . 206 . . . . . 206 . . . . . .36 . . . . . .30 . . . . . .30 . . . . . .40 . . . . . .40 . . . . . .93 . . . . . .97 . . . . . .93 . . . . . .97 .106, 163, 166 . . . . . 127 . . . . . 127 . . . . . 127 . . . . . 127 . . . . . 127 . . . . . .95 . . . . . .95 . . . . . 159 . . . . . .87 . . . . . 159 . . . . . .87 . . . . . 132 . . . . . 110 . . . . . .96 . . . . . 164 . . . . . 115 . . . . . 117 . . . . . 164 . . . . . .28 . . . . . 119 . . . . . 160 . . . . . 118 . . . . . 160 . . . . . 160 . . . 160, 206 . . . . . 160 . . . . . 160 . . . . . 160 . . . 160, 206 . . . . . 180 . . . . . 181 . . . . . .92 . . . . . . 4 . . . . . . 4 . . . . . .45 . . . . . .45 . . . . . .33 . . . . . .33 . . . . . .29 . . . . . 206 . . . . . .99 . . . . . .99 . . . 99, 206
Cat. no.
12659-017 12661-013 12662-011 12662-029 12676-011 12676-029 12677-019 12677-027 12678-017 12681-011 12681-011 12681-011 12681-011 12681-029 12682-019 12684-015 12684-031 12684015 . 12684023 . 12684031 . 12684049 . 12744-017 12744-017 12744-017 12746-012 12752-010 12753-018 12755-013 12760-013 12760-021 12760-021 12760013 . 12760021 . 12800-116 13018-015 13018-031 13150-016 13151-014 13256-029 13257-019 13290-010 13778-075 13778-150 14025-050 14025-076 14025-100 14025-126 14040-083 14040-091 14040-117 14040-174 14040-224 14065-049 14080-048 14155-048 14170-070 14170-088 14170-138 14170-195 14170-203 14175-053 14175-079 14175-129 14175-145 14185-045 14190-094 14190-094 14190-094 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . 175 . . . .96 . . . 190 . . . 190 . . . 189 . . . 189 . . . 109 . . . 109 . . . 109 . . . 180 . . . 181 . . . .89 . . . .92 . . . .89 . . . 175 . . . .88 . . . .88 . . . . 6 . . . . 6 . . . . 6 . . . . 6 . . . 180 . . . 181 . . . .92 . . . 118 . . . 114 . . . 114 . . . 114 . . . .88 . . . 119 . . . .88 . . . . 5 . . . . 5 . . . .30 . . . .45 . . . .45 . . . 158 . . . 158 . . . 125 . . . 116 . . . 116 . . . 133 . . . 133 . . . 140 . . . 140 . . . 140 . . . 140 . . . 138 . . . 138 . . . 138 . . . 138 . . . 138 . . . 140 . . . 139 . . . 139 . . . 140 . . . 140 . . . 140 . 140, 206 . 140, 206 . . . 140 . . . 140 . . . 140 . . . 140 . . . 140 . . . 118 . . . 119 . . . 138
246
Important Licensing Information: The products in this catalog may covered by one or more Limited Use Label Licenses (see the Invitrogen website www.invitrogen.com). By use of these products, you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for research use only, and are not intended for human or animal diagnostics, therapeutic, commercial, or other clinical uses unless otherwise specifically indicated in the applicable product Limited Use Label License(s).
Cat. no.
14190-136 14190-169 14190-326 14190-342 14190-359 14190-367 14200-067 14200-083 14287-072 14287-080 15023-021 15040-033 15070-063 15070-063 15090-046 15140-122 15140-122 15140-130 15140-148 15140-148 15140-163 15160-047 15160-070 15210-040 15212-040 15230-001 15230-071 15230-071 15230-089 15230-097 15230-147 15230-162 15230-196 15230-196 15230-204 15230-253 15230-261 15230-279 15230-788 15240-062 15240-062 15240-096 15240-096 15240-112 15250-061 15260-037 15290-018 15290-026 15338-100 15338-500 15400-054 15630-049 15630-056 15630-122 15640-055 15710-049 15710-049 15710-064 15710-072 15710-080 15750-045 15750-060 15750-078 15750-102 15918-014 15920-010 16000-036 16000-044 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . 138 . . . 138 . . . 138 . 138, 206 . 138, 206 . 138, 206 . . . 139 . . . 139 . . . 138 . . . 138 . . . 168 . . . 161 . . . 129 . . . 154 . . . 160 . . . 129 . . . 154 . . . 154 . . . 129 . . . 154 . . . 129 . . . 153 . . . 153 . . . 127 . . . 128 . . . 161 . . . 130 . . . 161 . . . 161 . . . 161 . . . 130 . . . 130 . . . 130 . . . 161 . . . 161 . 161, 206 . 161, 206 . . . 206 . . . 130 . . . 129 . . . 150 . . . 129 . . . 150 . . . 129 . . . 168 . . . 163 . . . 129 . . . 129 . . . 132 . . . 132 . . . 160 . . . 157 . . . 157 . . . 157 . . . 155 . . . 118 . . . 153 . . . 129 . . . 129 . . . 153 . . . 153 . . . 129 . . . 129 . . . 153 . . . 108 . . . 108 . . . 187 . . . 187
Cat. no.
16010-159 16010-167 16030-074 16050-122 16050-130 16070-096 16110-082 16120-099 16120-107 16140-063 16140-071 16141-061 16141-061 16141-079 16141-079 16170-078 16170-086 16210-072 16210-096 16250-078 16250-086 16447-100 16447-100 16447-100 16447-100 16447-500 16447-750 17001-074 17001-082 17005-042 17018-029 17018-029 17075-029 17100-017 17101-015 17104-019 17104-019 17105-041 17502-048 17502-048 17502-048 17503-012 17503-012 17504-044 17504-044 17701-038 17703-034 17704-024 17705-021 18045-054 18045-054 18045-070 18180-059 18200-048 18200-048 18300-012 18300-012 18324-012 18324-020 18324-111 20012-019 20012-043 20012-068 21012-026 21013-024 21013-024 21041-025 21051-040 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191 191 191 192 192 192 192 192 192 188 188 123 189 123 189 191 191 192 192 190 190 132 180 181 .92 132 132 127 127 110 130 158 161 158 158 119 158 158 115 117 165 115 164 115 164 113 113 113 113 123 .29 123 167 103 167 103 167 133 133 133 141 141 141 .99 .30 .32 .33 149
Cat. no.
21056-023 21060-017 21060-017 21063-029 21083-027 21090-022 21090-055 21103-049 21103-049 21127-022 21180-021 21300-017 21300-058 21340-054 21340-054 21430-020 21530-027 21540-026 21575-022 21600-010 21600-044 21600-069 21700-018 21700-026 21710-025 21720-024 21720-024 21765-029 21765-037 21810-031 21850-029 21875-034 21875-042 21875-091 21875-158 21885-025 21885-108 21969-035 21980-032 21980-065 21985-023 22320-022 22330-021 22340-020 22350-029 22390-025 22400-147 22400-154 22551-022 22561-021 22561-054 22571-020 22571-038 22600-050 22600-134 22600-134 23017-015 23017-015 23181-010 24020-091 24040-032 24040-032 24040-032 24040-032 24040-032 25030-024 25030-032 25030-081 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36 107 166 .30 .37 .41 .41 114 117 .34 .38 139 139 107 .42 .41 .29 .29 .42 139 139 139 .34 .34 .35 100 .46 .34 .34 154 155 .45 .45 .45 .45 .32 .32 .30 .36 .36 168 .32 .37 .38 .38 .34 206 206 .46 .40 .40 .40 .40 .95 .43 .95 162 . 6 119 140 103 167 180 181 .92 118 149 130
Cat. no.
25030-081 25030-081 25030-081 25030-123 25030-123 25030-149 25050-014 25050-030 25080-060 25080-060 25080-102 25200-056 25200-072 25200-114 25300-054 25300-062 25300-096 25300-104 26010-066 26010-074 26050-070 26050-088 26140-079 26140-087 26170-035 26170-043 26250-084 26400-036 26400-044 26600-023 26600-080 27016-021 27016-021 27250-018 31035-025 31035-025 31053-028 31095-029 31095-052 31100-019 31100-027 31150-022 31150-030 31220-023 31330-038 31330-095 31331-028 31331-028 31331-093 31331-093 31350-010 31550-023 31600-083 31600-091 31765-027 31765-068 31870-025 31870-074 31885-023 31885-049 31966-021 31966-047 31980-022 31980-048 31985-047 31985-047 31985-054 31985-054 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 180 181 .92 130 149 130 160 160 130 157 130 160 160 160 160 160 160 160 191 191 192 192 188 188 191 191 192 189 189 .37 .37 130 .45 161 112 117 .30 .40 .40 .38 .38 .38 .38 .46 .33 .33 120 .33 120 .33 119 .34 .32 .32 .34 .34 .45 .45 .32 .32 .31 .31 .36 .36 .43 .94 .43 .94
Cat. no.
31985-062 31985-062 32360-026 32430-027 32430-100 32561-029 32561-094 32571-028 32571-093 33016-015 33016-015 35050-038 35050-038 35050-061 35050-079 35050-087 35050-087 37010-022 39-8401 . 402-03D . 41-1000 . 41-1100 . 41-1200 . 41-4000 . 41-4400 . 41010-026 41061-029 41090-028 41090-093 41300-021 41300-070 41400-045 41500-018 41500-083 41965-039 41965-047 41965-062 41965-120 41966-029 41966-052 42200-014 42200-030 42360-024 42360-081 42430-025 42430-082 51200-046 51300-044 51500-056 51800-019 51800-035 51800-043 51985-026 51985-026 51985-042 51985-042 52100-039 52400-017 52400-025 52400-041 61100-087 61100-103 61870-010 61870-044 61870-143 61870-150 61965-026 61965-059 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43 .94 .41 .31 .31 .41 .41 .41 .41 125 162 123 149 130 130 123 149 110 118 . 3 119 119 119 119 119 .28 .40 .41 .41 .37 .37 164 .40 .40 .30 .30 .30 .30 .30 .30 .36 .36 .41 .41 .30 .30 .41 164 165 .45 .45 .45 .43 .94 .43 .94 .30 .45 .45 .45 .40 .40 .45 .45 206 206 .31 .31
Catalogue Number Index 247
www.invitrogen.com
Catalog Number Index

Cat. no.
70011-036 70011-051 70013-016 70013-065 72400-021 72400-054 72400-146 72400-153 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . 141 141 141 141 .45 .45 206 206
Cat. no.
CTP2111 CTP2113 CTP3011 CTP3013 CTP9211 CTP9213 CTP9411 CTP9413 DGCP10 DGCP20 DGCS10 DGCS20 HMCPIL HMCPIS HMCPML HMCPMS HMCPTL HMCPTS HMCS1L HMCS1S HMCS2L HMCS2S HMCS3L HMCS3S HMCSTL HMCSTS K4930-00 K4945-00 K4950-00 K4960-00 K4965-00 K4980-00 K4990-00 K5910-00 K8300-01 K9000-01 K9000-20 MKCP10 MKCP20 MKCS10 MKCS20 MSCP10 MSCP20 MSCS10 MSCS20 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
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Cat. no.
PHC0061 . PHC0063 . PHC0064 . PHC0065 . PHC0066 . PHC0071 . PHC0073 . PHC0075 . PHC0076 . PHC0084 . PHC0101 . PHC0103 . PHC0104 . PHC0105 . PHC0115 . PHC0131 . PHC0134 . PHC0135 . PHC0174 . PHC0186 . PHC0191 . PHC0194 . PHC0201 . PHC0204 . PHC0205 . PHC0211 . PHC0213 . PHC0214 . PHC0215 . PHC0811 . PHC0813 . PHC0814 . PHC0815 . PHC0816 . PHC0881 . PHC0883 . PHC0884 . PHC0885 . PHC1011 . PHC1014 . PHC1015 . PHC1031L PHC1033L PHC1034Z PHC1051 . PHC1053 . PHC1054 . PHC1055 . PHC1061 . PHC1063 . PHC1065 . PHC1066 . PHC1074 . PHC1081 . PHC1083 . PHC1084 . PHC1085 . PHC1094 . PHC1095 . PHC1101 . PHC1104 . PHC1111 . PHC1114 . PHC1115 . PHC1116 . PHC1123 . PHC1124 . PHC1154 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
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Cat. no.
PHC1174 . PHC1204 . PHC1234 . PHC1244 . PHC1254 . PHC1264 . PHC1331 . PHC1333 . PHC1335 . PHC1336 . PHC1341 . PHC1343 . PHC1344 . PHC1346 . PHC1364 . PHC1431 . PHC1433 . PHC1434 . PHC1471 . PHC1473 . PHC1474 . PHC1506 . PHC1514 . PHC1524 . PHC1536 . PHC1544 . PHC1564 . PHC1574 . PHC1585 . PHC1594 . PHC1601 . PHC1603 . PHC1604 . PHC1615 . PHC1624 . PHC1625 . PHC1634 . PHC1674 . PHC1684 . PHC1691 . PHC1694 . PHC1804 . PHC2011 . PHC2013 . PHC2015 . PHC2031 . PHC2033 . PHC2035 . PHC2054 . PHC2111 . PHC2113 . PHC2115 . PHC2116 . PHC3011 . PHC3013 . PHC3015 . PHC3015L PHC3016 . PHC4014 . PHC4031 . PHC4033 . PHC4044 . PHC4054 . PHC4064 . PHC4244 . PHC4814 . PHC5015 . PHC7015 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
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Cat. no.
PHC7024 PHC7041 PHC7043 PHC7044 PHC7045 PHC7056 PHC7064 PHC7074 PHC7075 PHC7084 PHC7104 PHC7141 PHC7143 PHC7145 PHC7146 PHC7204 PHC7914 PHC8011 PHC8013 PHC8014 PHC8015 PHC8021 PHC8023 PHC8024 PHC9151 PHC9154 PHC9171 PHC9173 PHC9174 PHC9211 PHC9213 PHC9214 PHC9215 PHC9221 PHC9224 PHC9241 PHC9243 PHC9244 PHC9251 PHC9253 PHC9254 PHC9255 PHC9261 PHC9264 PHC9271 PHC9274 PHC9281 PHC9283 PHC9284 PHC9285 PHC9291 PHC9293 PHC9294 PHC9295 PHC9311 PHC9313 PHC9314 PHC9315 PHC9341 PHC9343 PHC9344 PHC9345 PHC9381 PHC9383 PHC9384 PHC9385 PHC9391 PHC9393 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
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AB
A1000701 A10142-01 A1014201 A10234-01 A10240-01 A10241-01 A1036901 A1036902 A1048001 A1048002 A1048301 A1048301 A10485-01 A1049201 A1064401 A1091501 A1098201 A1098202 A1099201 A1099202 A1099901 A1100-001 A1113-802 A1113-803 A1113-902 A1113-903 A1134001 A1134301 A1142801 A1142802 A1142803 A12856-01 A12858-01 A12859-01 A12860-01 A12861-01 A1287-301 A1287-302 A1287-303 A1287-304 A13261 . . A13262 . . B855-02 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 . 4 . 4 .91 .91 .91 156 156 . 5 . 5 . 5 .88 . 3 149 . 5 . 6 . 6 . 6 . 4 . 4 .92 .91 151 151 151 151 . 6 . 5 . 5 . 5 . 5 3, 4 3, 4 . 4 . 4 . 4 161 161 161 161 171 171 174
P
PHB0044 PHB0064 PHC0011 PHC0013 PHC0015 PHC0017 PHC0021 PHC0023 PHC0026 PHC0027 PHC0031 PHC0033 PHC0034 PHC0035 PHC0041 PHC0043 PHC0044 PHC0045 PHC0052 PHC0055 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15 .15 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .16 .15 .16
CM
CTP0021 CTP0023 CTP0041 CTP0043 CTP0061 CTP0063 CTP0071 CTP0073 CTP0261 CTP0263 CTP2011 CTP2013 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 3 3 3 4 4 3 3 4 4 3 3
248
Cat. no.
PHC9394 . PHC9411 . PHC9413 . PHC9414 . PHC9415 . PHC9431 . PHC9434 . PHC9461 . PHC9461 . PHC9464 . PHC9464 . PHC9471 . PHC9474 . PHC9501 . PHC9504 . PHC9511 . PHC9513 . PHC9514 . PHC9531 . PHC9533 . PHC9534 . PHC9541 . PHC9543 . PHC9544 . PHC9551 . PHC9553 . PHC9554 . PHC9561 . PHC9563 . PHC9564 . PHE0011 . PHE0023 . PHG0014 . PHG0021 . PHG0021 . PHG0021L PHG0021L PHG0023 . PHG0023 . PHG0024 . PHG0024 . PHG0026 . PHG0026 . PHG0035 . PHG0041 . PHG0043 . PHG0044 . PHG0045 . PHG0046 . PHG0051 . PHG0094 . PHG0126 . PHG0134 . PHG0154 . PHG0174 . PHG0184 . PHG0194 . PHG0204 . PHG0214 . PHG0224 . PHG0234 . PHG0244 . PHG0254 . PHG0261 . PHG0261 . PHG0261 . PHG0263 . PHG0263 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
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Cat. no.
PHG0264 . PHG0264 . PHG0266 . PHG0266 . PHG0271 . PHG0273 . PHG0274 . PHG0276 . PHG0286 . PHG0296 . PHG0311 . PHG0311 . PHG0311L PHG0311L PHG0313 . PHG0314 . PHG0314 . PHG0315 . PHG0315 . PHG9014 . PHG9074 . PHG9074 . PHG9114 . PHG9202 . PHG9203 . PHG9204 . PHG9211 . PHG9214 . PHG9302 . PHG9305 . PHP0024 . PHP0025 . PHP0034 . PHR3015 . PHR3025 . PHR5001 . PHS0044 . PHZ0014 . PMC4051 . PMC4054 . PMG0041 . PMG0043 . PMG0044 . PMG0045 . PRC0011 . PRC0014 . PRC0021 . PRC0024 . PRC0031 . PRC0034 . PRC0051 . PRC0053 . PRC0054 . PRC0061 . PRC0063 . PRC0064 . PRC0104 . PRC0131 . PRC0133 . PRC0134 . PRC0181 . PRC0184 . PRC0811 . PRC0813 . PRC0814 . PRC0815 . PRC1011 . PRC1014 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12 .15 .12 .15 .15 .15 .15 .15 .15 .17 124 .15 124 .15 .15 124 .15 124 .15 .15 124 .16 .17 .17 .17 .17 .17 .17 .17 .17 .15 .15 .17 .17 .17 .15 .15 .15 .13 .13 124 124 124 124 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18
Cat. no.
PRC1015 PRC1054 PRC1066 PRC1071 PRC1074 PRC2011 PRC2015 PRC2016 PRC3013 PRC3014 PRC3015 PRC4014 PRC4024 PRC4033 PRC4034 PRC4035 PRC7015 PRG0111 PRG0113 PRG0114 PRG0115 PSC0011 PSC0014 PSC0021 PSC0024 PSC0041 PSC0044 PSC0045 PSC0061 PSC0064 PSC0101 PSC0104 PSC0151 PSC0154 PSC0881 PSC0884 PSC2011 PSC2014 PSC2015 PSC3011 PSC3014 PSC4031 PSC4034 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Page
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .18 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19 .19
Cat. no.
Page
Cat. no.
Page
Catalogue Number Index
RV
R210-01 . R210-01 . R250-01 . R250-05 . R705-07 . R7799-105 R790-07 . RBCP10 . . RBCP20 . . RBCS10 . . RBCS20 . . RTCS10 . . SA100-04 . TRCS10 . . TRCS20 . . TRCS30 . . V492-20 . V494-20 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 151 155 155 .97 119 .94 183 183 183 183 183 108 183 183 183 .97 .97
-NGF Recombinant Human is this still in catalogue?
249
www.invitrogen.com
Notes:
250
gibco Cell Culture Catalogue
Notes:
Catalog Number Index 251

www.invitrogen.com
Notes:
252
Notes:
Catalogue Number Index 253

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Notes:
254

14 - 1 Cultivo Cel Animales Gibco Catalogue Edition 49

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14 - 1 Cultivo Cel Animales Gibco Catalogue Edition 49

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gibco cell culture catalogue

Resources web links

Cell Culture Products

Containers and closures

Production and Preparation

Custom Cell Culture Products

Custom liquid medium concentrates

Advanced Granulation Technology (AGT)

gibco cell culture catalogue

Powdered Cell Culture Products

Ordering and Technical Support

Order by Phone, Mail, or Fax

gibco cell culture catalogue

gibco cell culture catalogue

Chapter 2 Growth factors, Cytokines and Chemokines with high bioactivity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Chapter 3 Basal Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Chapter 4 Serum-free and Specialty Media and Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Chapter 5 Balanced Salt Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .137

Chapter 6 Standard Cell Culture Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .147

gibco cell culture catalogue

Chapter 7 Eukaryotic Cell Lines

Chapter 8 Animal Sera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .185

Gibco Cell Therapy Systems (CTS)

gibco cell culture catalogue

Gibco CTS Immunotherapy products

AIM V Medium CTSTherapeutic-grade, serum-free cell expansion medium

CTS cytokines and growth factors

OpTmizer CTS T-Cell Expansion SFM

Gibco CTS products for stem cells

gibco cell culture catalogue

Gibco Extracellular Matrices

Bovine and Rat Tail Collagen I

AlgiMatrix 3D Culture System

Natural Mouse Laminin

gibco cell culture catalogue

Growth factors, Cytokines and Chapter 2 Chemokines with high bioactivity

Chapter 2 Growth factors, Cytokines, and Chemokines with high bioactivity

Growth factors, Cytokines and Chemokines with high bioactivity

High Biological Activity

Multiple Packaging Sizes

How are your cytokines, chemokines, and growth factors manufactured?

Carrier-free Sterile-filtered prior to

Expressed in a host system that yields a biologically active protein

Purified to produce a highly pure, homogenous solution of active protein

formulated and lyophilized to ensure maximum stability

gibco cell culture catalogue

How is the quality of your growth factors ensured?

2 Growth factors, Cytokines and Chemokines with high bioactivity

2.2 2.0 1.8

ED50 (ng/mL) GIBCO = 0.26 Competitor = 0.58

280,000 240,000 200,000

1.6 1.4 1.2

160,000 120,000 80,000 40,000 0 0

1.0 0.8 0.001 0.01 0.1 1.0 10 100

1.4 290 300

1,000 2,000 3,000 4,000 5,000 6,000 7,000

Chapter 2 Growth factors, Cytokines, and Chemokines with high bioactivity

Gibco Recombinant human FGF-basic

gibco cell culture catalogue

Gibco LIF Recombinant

Experiment#1 Recombinant LIF

2.3 2.1 1.9 1.7 1e-4