The stable tetrazolium salt WST-1 is cleaved to a soluble formazan by a complex cellular mechanism that occurs primarily at the cell surface. This bio-reduction is largely dependent on the glycolytic production of NAD(P)H in viable cells. Therefore, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture. Cells grown in a 96-well tissue culture plate are incubated with the WST-1 reagent for 0.5 – 4 hours. After this incubation period, the formazan dye formed is quantitated with 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 184.108.40.206) which belongs to the respiratory chain of the mitochondria, and is only active in metabolically intact cells.
The Cell Proliferation Reagent WST-1 is used for the nonradioactive, spectrophotometric quantification of cell proliferation and viability in cell populations using the 96-well-plate format. Measurement of cell proliferation in response to growth factors, cytokines, mitogens, and nutrients.
- Analysis of cytotoxic and cytostatic compounds, such as anti-cancer drugs and other pharmaceutical compounds
- Assessment of growth inhibitory antibodies and physiological mediators.
Working concentration: For best results, add 10 μl/well Cell Proliferation Reagent WST-1 to the cells already cultured in 100 μl/well (1:10 final dilution). Using the 100 μl/well cell culture volume, one vial will be sufficient to perform 2500 tests (25 microplates). Note: If the cells are cultured in 200 μl/well, add 20 μl/well Cell Proliferation Reagent WST-1.
The Cell Proliferation Reagent WST-1 is shipped on dry ice and should be stored at -15 to -25°C, protected from light. Once thawed it can be stored at 2-8°C, if protected from light, for several weeks. For longer storage it is recommended to store the Cell Proliferation Reagent WST-1 in aliquots at -15 to -25°C.
Celltechgen provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please request the Safety Data Sheet (SDS) provided for the product.