Member since: 2022
Organization: University for Applied Sciences Coburg
Application Area: Toxicity Tests
"The Kit was easy to use but lacked some visible information about additional necessary chemicals. It would have been nice to see on the product's website which other chemicals are mandatory to utilize the Kit instead of reading it in the protocol. Otherwise, we had no problems. The protocol came with helpful, theoretical background of the reaction and was easy to understand."
Member since: 2020
Excellent kit! Makes my work easier.
Application Area:Measurement of cell proliferation, analysis of cytotoxic and cytostatic compounds
"Its use is really safe and easy. The kit eliminates radioactive isotopes, washing steps, and additional reagents. Its accuracy is remarkable, the absorbance obtained strongly correlates to the cell number. Its high sensitivity detects low cell numbers, and it’s fast since it processes a large number of samples using a multi-well ELISA reader."
The Cell Proliferation Kit I (MTT) is a colorimetric assay for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity. Sample material is either adherent or suspension cells cultured in 96-well microplates.
Colorimetric assays analyze the number of viable cells by the cleavage of tetrazolium salts added to the culture medium. This technique requires neither washing nor harvesting of cells, and the complete assay, from microculture to data analysis by an ELISA reader, is performed in the same microplate.
MTT was the first tetrazolium salt described. It is cleaved to formazan by enzymes of the endoplasmic reticulum. This bioreduction occurs in viable cells only, and is related to NAD(P)H production through glycolysis. Therefore, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture.
Colorimetric assay (MTT based) for the nonradioactive quantification of cellular proliferation, viability, and cytotoxicity.
Features and Benefits
The assay is based on the cleavage of the tetrazolium salt MTT in the presence of an electron-coupling reagent. The water-insoluble formazan salt produced must be solubilized in an additional step. Cells grown in a 96-well tissue culture plate are incubated with the MTT solution for approximately 4 hours. After this incubation period, a water-insoluble formazan dye is formed. After solubilization, the formazan dye is quantitated using a scanning multi-well spectrophotometer (ELISA reader). The measured absorbance directly correlates to the number of viable cells.
Cell proliferation and viability assays are of particular importance for routine applications in cell biology. Tetrazolium salts (e.g., MTT, XTT, WST-1) are particularly useful for this type of analysis. Tetrazolium salts are cleaved to formazan by the succinate-tetrazolium reductase system (EC 220.127.116.11) which belongs to the respiratory chain of the mitochondria, and is only active in metabolically intact cells.