A) Serum withdrawal prior to treatment / lower the serum concentration from 10% to 0.5 or 2.5%
OR
B) No serum withdrawal at all prior to treatment and perform treatment in regular media.
If your cells can tolerate it, you'd probably see a larger activation (relative to the untreated) with a serum starvation in low serum media. In the presence of serum, you could possibly have some level of activation of your desired protein due to the presence of a myriad of growth factors. That said, some proteins are activated so robustly by growth factor treatment that it doesn't matter which treatment protocol you use.
I also agree with the Dr Mullenbrock. However, the protocol also depends on your cell type.
Above answers can be followed, for my experiments I always like to starve cells overnight in presence of 0.2%FBS............always works for my experiments..............especially for Growth factors...................good luck
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