WST-1 is a cytotoxic, water-soluble tetrazolium salt that induces intracellular mitochondrial dehydrogenases to undergo NADH-dependent enzymatic reactions, releasing water-soluble methylbenzene products.WST-1 can be used to detect cell proliferation and cellular activity by assaying the uptake values, and can be used to determine superoxide dismutase activity in the WST-1 plate assay.

Instructions
1. Preparation of WST-1 solution: WST-1 powder is fully dissolved in DMSO to prepare a 10 mg/mL WST-1 stock solution. The prepared solution should be stored away from light, can be stored at 4℃ for one week, and can be stored at -20℃ for half a year, and avoid repeated freezing and thawing.
2. Cell inoculation: According to the purpose of the experiment, add an appropriate amount of cell suspension to a 96-well plate and culture overnight to ensure cell adhesion.
Treat cells: According to the needs of the experiment, add different concentrations of the drug or stimulant to be tested to each well and continue to culture for an appropriate time.
3. Add WST-1 reagent: Add 5-10 μM WST-1 working solution to each well and mix gently.
4. Incubation: Place the culture plate in a cell culture incubator and incubate for 0.5 to 4 hours. The specific incubation time can be optimized according to the cell type and density, usually 1-2 hours.
5. Determine absorbance: Use a microplate reader to measure the absorbance of each well at a wavelength of 450nm and calibrate it with a reference wavelength of 600nm. The level of absorbance reflects the proliferation or survival of cells.