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CellTiter 96 Non-Radioactive Cell Proliferation Assay

by Promega
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  • Overview
  • Protocols
  • Specifications
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  • The CellTiter 96® Non-radioactive Cell Proliferation 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 and 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 With 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.

    Convenient and Adapts to Your Needs: Requires no weighing or mixing of dye compounds and follows either a 4-hour or overnight protocol.

  • Protocols

    Complete Protocol

    Download PDF

    CellTiter 96® Non-Radioactive Cell Proliferation Assay Technical Bulletin

    PDF (874 KB)

    Quick Protocols

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    CellTiter 96 Non-Radioactive Cell Proliferation Assay Quick Protocol FB045

    PDF (101 KB)

  • Certificate of Analysis

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    Storage Conditions


    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

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