MTT-Assay: Cell Viability [Mosmann 1983, La Fontaine et al 1998, Southon et al

2004] modiefied
-

5/10/2005

add MTT solution [5 mg/mL] in an amount equal to 10% of the cell culture
volume

incubate at appropriate temperature for the used cell-type for 1-4 hours

add acetic isopropanol

a) if cells are attached firmly to cell culture vessle, remove growth medium
and add same amount of acetic isopropanol
b) if cells are loosely attached to cell culture vessle, add acetic isoropanol
directly to the culture in an amount equal to the original culture volume

within 1 hour (all crystals should be dissolved by then, otherwise reduce

incubation time with MTT) after adding acetic isopropanol read the absorbance
at 570 nm and subtract background absorbance read at 690 nm

MTT-solution:
-

5 mg/mL Thiazolyl Blue Tetrazolium Bromide (MTT; Sigma, M2128) in 1xPBS

(pH 7.4)

sterile filter to remove crystals and to sterilize

store at 20 C (development of dark color or formation of crystals indicate

deterioation)

Acetic Isopropanol:
0.1 N HCl in isopropanol
References
La Fontaine S, Firth SD, Lockhart PJ, Brooks H, Parton RG, Camakaris J, Mercer JF
(1998), Functional analysis and intracellular localization of the human menkes protein
(MNK) stably expressed from a cDNA construct in Chinese hamster ovary cells
(CHO-K1), Hum.Mol.Genet. 7: 1293-1300
Mosmann T (1983), Rapid colorimetric assay for cellular growth and survival:
application to proliferation and cytotoxicity assays, J.Immunol.Methods 65: 55-63
Southon A, Burke R, Norgate M, Batterham P, Camakaris J (2004), Copper
homoeostasis in Drosophila melanogaster S2 cells, Biochem.J. 383: 303-309