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Cytotoxicity assay using WST-1

AR Asmaa Reda
AR Alaa Refaat
AA Ahmed A. Abd-Rabou
AM Ali Mokhtar Mahmoud
MA Mohamed Adel
SS Salwa Sabet
SA Sameh Saad Ali
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Cells were seeded in 96-well plates at a density of 1 × 104 cells/well and allowed to settle overnight in 5% CO2 at 37 °C. The medium was discarded and was replaced with a fresh one containing different final concentrations of iodoacetate (0–50 μM) in triplicates and incubation was continued for 4 h and 24 h. At the end of the incubation period, the cells were rinsed with PBS and treated with 10 μl of the cell proliferation assay reagent WST-1 (Sigma-Aldrich) for 4 h. The amount of formazan dye formed as a result of the cleavage of the stable tetrazolium salt WST-1 was measured spectrophotometrically at λ = 450 nm using a FLUOstar Omega microplate reader (BMG Labtech, Germany) and is directly correlated to the cytotoxic effect of the drug.

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