I've worked with p38MAPK inhibitors before, and those work fine with thirty minute treatments. I'm now working with a GSK3 inhibitor, and in literature, people have mostly treated cells overnight, but those end points were apoptosis or neurite outgrowth, both of which would take a long time to attain. Since my end point is just to look at tau phosphorylation, I am wondering if I can get away with a shorter treatment with ten micromolar of this cell-permeable inhibitor.
Most or the studies are done using SB inhibitors at 5uM for 24-48 h in primary neurons. Increasing the concentration and to shorten the treatment time is a subject of optimization: you don't want to have "off target" effect of a high concentration on cell viability. To generate a time/concentration curve looking for tau phosphorylation should be a straight forward experiment.
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