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The CellTiter 96 Assay is a collection of qualified reagents that provide a convenient method of determining viable cell number. A modification of the MTT assay method described by Mosmann, the assay incorporates several improvements to the method that address previous technical problems including: 1) serum protein precipitation caused by adding organic solvent; 2) interference by phenol red; 3) incomplete solubilization of the formazan crystals resulting in lower sensitivity; and 4) stability of the colored product. The assay is performed by adding a premixed, optimized Dye Solution to culture wells of a 96-well plate, usually containing various concentrations of growth factor or test substance. During a 4-hour incubation, living cells convert the MTT tetrazolium component of the Dye Solution into a formazan product. The Solubilization/Stop Solution is added to the culture wells to solubilize the formazan product, and the absorbance at 570nm is recorded using a 96-well plate reader.
