I am testing effect of drug on expression of five proteins. I guess I shall use five wells. Each for one protein and the sixth well for control with no drug. I don't know how many cells to seed for two days treatment. Thermo fisher density table says 0.3 million cell for six well plate.
Will you be including biological replicates by combining runs? Ideally you would probe for 1 target + 1 control/normalizer/reference with all biological replicates rather than 5 targets 1 control in a single biological replicate. For cell numbers see below:
https://www.researchgate.net/post/Minimal_amount_of_cells_protein_for_Western_Blot
Thanks a lot. I think I will try 100.000 in one plate and 300.000 in one plate and wait two days to see cell confluency. Is cell confluency at last day of treatment should be 80% same as proliferation assay? I measure protein expression after adding drug.
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