Oxidative Stress Assay in Cancer Cells

SKOV-3 and OVCAR-3 cells were seeded as monolayers at 20,000 cells per well in a black 96-well plate and cultured overnight. Following removal of media, cells were washed once with PBS, then incubated for 1 h with the Cellular Reactive Oxygen Species Detection reagent (Red Fluorescence, Abcam, UK, 186027). An IC20 concentration of selenite or SeNPs (mentioned previously) was then added and the plate was incubated at 37 °C for the duration of the assay. Fluorescence was analysed at different time points from 30 min to 10 h (excitation filter 520 nm, emission filter 605 nm, BMG Labtech Fluostar Omega, UK).