I want to measure the proliferation rate of 293T Flp in T-reX cells after the induction of the expression of my protein with Tetracycline.
I should add the MTT to the cells after 24-48-72-96 hrs, but at the same time I have to change the medium with fresh tetracycline every 24hrs. Does this change of the medium affect the reliability of the assay?
If I were you, this is what I would do. In a 96 well plate, add cells to four different columns ( first column for 24 hrs, second for 48 hrs and so on till the 96th hour). Add your drug to the columns and after 24 hours add MTT to the column that is designated for 24th hour reading. Let that column of cells incubate with MTT for 1 or 2 hours and take a reading using a plate reader. You can change the media in the other columns that don't have MTT with fresh Tetracycline. And after 48 hours you can MTT to the 48 hour column. I hope this helps.
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