Dr. Liang, have a look at my paper
"Assessment of the ability of seaweed extracts to protect against hydrogen peroxide and tert-butyl hydroperoxide induced cellular damage in Caco-2 cells.". I worked with tert-butyl hydroperoxide in this trial. I would recommend using a dose response from 0 to 2000 micro molar. I'd test cell viability first using neutral red and MTT assays. From here you can decide what concentrations to proceed with.
For MTT and neutral red assay we choose the maximum concentration that could be added to cells without affecting cell viability.
To find the ideal concentrations of tert-butyl hydroperoxide to induce oxidative stress I used a dose response (below the concentration that affected cell viability in the MTT and Neutral Red assays).
I think they discontinued the product I used. Try the following product instead: Sigma Aldrich Luperox® TBH70X, tert-Butyl hydroperoxide solution (product code: 458139).
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