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#MTT assay

2 public questions tagged with this topic.

Animal Tissue Culture 3

The purple color in MTT assay is due to formation of:

Purple color appearing in wells after MTT incubation arises from formation of formazan product 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan. Enzymatic reduction transfers electrons from succinate via succinate dehydrogenase, NADH and NADPH to tetrazolium ring breaking positively charged quaternary nitrogen and producing neutral formazan with extensive conjugated double bond system delocalizing electrons, absorbing green-yellow wavelengths around 570 nm and appearing purple. Crystals observable microscopically as dark needle-like precipitates clustered around nucleus and in cytoplasm befor

Ref: Berridge et al Arch Biochem Biophys 2005 MTT formazan formation mechanism; Abcam MTT assay protocol formazan purple product succinate dehydrogenase.

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Animal Tissue Culture 3

MTT assay measures:

MTT assay developed by Tim Mosmann in 1983 measures cell viability and proliferation based on reduction capacity of living cells. Principle uses yellow tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide which enters cells via endocytosis and is reduced by mitochondrial succinate dehydrogenase of complex II and cytosolic NAD(P)H-dependent oxidoreductases cleaving tetrazolium ring with electrons from NADH and NADPH producing insoluble purple formazan crystals accumulating intracellularly in mitochondria and lipid droplets. Metabolically incompetent dead cells fail to r

Ref: Mosmann T J Immunol Methods 1983 MTT viability assay; ATCC MTT Cell Proliferation Assay mitochondrial dehydrogenase reduction protocol.

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