For exemple 3 hours of drug and then 3 hours of MTT. Can I say the total drug treatment was 6 hours?
Hello Francesca Valenti
No, it is incorrect.
This is because MTT reagent is not the part of treatment. It evaluates the cell viability after the drug treatment in the form of the quantity of formazan (presumably directly proportional to the number of viable cells) which is measured by recording changes in absorbance at 570 nm using a plate reading spectrophotometer. A reference wavelength of 630 nm is sometimes used, but not necessary for most assay conditions.
After having treated the cells with the drug, the drug containing media is removed and MTT reagent is added to the wells. Instead, I would consider the total time for MTT assay as 6 hours consisting of 3 hours of drug treatment and then 3 hours of MTT reagent incubation time.
Best.
Why did you use a 3 h treatment of cells by MTT ? In my experience, 30 minutes incubation with MTT is sufficient to get a robust answer and therefore you have to consider only your time of treatment of 3 hours as mentionned by Malcolm
thank you, many protocols say to incubate MTT for 2-4 hours. I will try the 30 minutes time in any case.
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