CellTiter 96® Non-Radioactive Cell Proliferation Assay (MTT)

The CellTiter 96® Assay is a collection of qualified reagents that provide a convenient method of determining viable cell number. The CellTiter 96® Assay is a modification of the MTT assay method described by Mosmann and 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 CellTiter 96® Assay is performed by adding a premixed, optimized Dye S...

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The CellTiter 96® Assay is a collection of qualified reagents that provide a convenient method of determining viable cell number. The CellTiter 96® Assay is a modification of the MTT assay method described by Mosmann and 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 CellTiter 96® 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. If you currently use a [³H]-thymidine incorporation assay, the addition of Dye Solution can be substituted for the pulse of radioactive thymidine at the time point in the assay when the pulse of [³H]-thymidine is usually added. The Solubilization/Stop Solution is then added to the culture wells to solubilize the formazan product, and the absorbance at 570nm is recorded using a 96-well plate reader. In addition, direct comparison between [³H]-thymidine incorporation and tetrazolium conversion have demonstrated less than a 5% difference between the two assays for determination of growth factor content of several samples.

Features - Benefits

• Gain Sensitivity: Detect as few as 1,000 cells/well with a 96-well plate reader. Greater sensitivity than the neutral red assay procedure.
• Use a Variety of Cells: Assay mammalian, plant and yeast cells.
• Non-Radioactive: Requires no scintillation cocktail or radioactive waste disposal.
• Save Time: Perform the assay in a 96-well plate with no washing steps, no cell harvesting and no scintillation counting.
• Adapt to Your Needs: Follow either a 4-hour or overnight protocol.
• Convenient: Requires no weighing or mixing of dye components.

Applications

• Cell proliferation.
• Cell viability.
• Cytotoxicity.
• Cell attachment.
• Chemotaxis.

For more information, see the Protocols & Applications Guide.

References

1. Mosmann, T. (1983) J. Immunol. Meth. 65, 55–63.
2. Tada, H. et al. (1986) J. Immunol. Meth. 93, 157–65.

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