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TABLE 2 CHEMICAL PROPERTIES OF COMMONLY USED DETERGENTS Ionic Detergents SDS CHO DOC C16 + + Dialyzable + Ion exchangeable + Complexes ions Strong A280 Assay Interference + Cold Precipitates High Cost + Availability Toxicity + Ease of Purification + Radiolabeled Defined Composition Auto-oxidation + + + + + + + + + + + + + + + + + + + + + + + + + +/+ + + + + + + + +/+ + +/+ + + + + + + + + + + + +/+ +/+ +/+/+ + + + + + + + + Nonionic Detergents LYS CHA ZWI OGL DIG C12 LUB TNX NP40 T80 +/+ +/+/+ -
Choice of Detergents Ionic detergents are very good solubilizing agents, but they tend to denature proteins by destroying native threedimensional structures. This denaturing ability is useful for SDS-PAGE but is detrimental where native structure is important for functional activity. It should be noted that antibodies usually retain their binding activity at 0.1% SDS or less. Nonionic and mildly ionic detergents are less denaturing and can often be used to solubilize membrane proteins while retaining protein-protein interactions. The following detergents have detrimental properties for some procedures: 1. Phenol containing detergents (ie., Triton X-100 and NP40) have a high absorbance at 280nm and thus interfere with protein monitoring at that OD. Lubrol may be substituted. Phenol-containing detergents also induce precipitation in the Folin (Lowry) protein assay. They can also be iodinated...and thus should not be used if iodinating proteins.
2. High micellar molecular weight interferes with gel filtration and not easily removed by dialysis. 3. Sodium cholate and sodium deoxycholate are insoluble below pH 7.5. Above an ionic strength of 0.1%SDS they will often crystalize. 4. Ionic detergents interfere with nondenaturing electrophoresis and isoelectric focusing.