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Timing for cyclinD1 and c-myc - (Jul/15/2008 )

Hi all,
I am going to start doing westerns. I will treat the cell line and then will check phospho- and total Erk 1/2, cyclin D1, c-myc levels in drugged vs. control wells. For phospho Erk 1/2 as far as I have searched the literature I need to do the treatment for around 30 minutes and then lyse the cells. Cyclin D1 and c-myc are early G1 phase proteins. But there are very different timing points for these in the literature. Could anyone recommend me a good time point to catch phosphorylation of these proteins? Is 24 hours too long? Many thanks in advance,


man! these things differs between cell way for you is to a time course experiment, if WB is too much you can also find the same by FACS.