yes, I used anti-phospho p38 for immunoprecipitation, because it was in the p38 MAP kinase-assay kit from NEB.
No, I do not know whether the anti-phospho ATF-2 antibody is working. Do you know a positiv control to test this?
For cell lysis I used RIPA-buffer (containing NP-40 and sodium deoxycholate)
and dilute the sample in NEB-lysis buffer (containing Triton X-100) prior
may be I will try the agarose-conjugated anti-p38 antibody from Santa Cruz. Which antibody do you use? sc-535 AG or sc-728 AG???
Do you perform a pre-clearing step before immunoprecipitation? I found, that pre-clearing with Protein A-agarose beads "precipitated" the p38!!!
Then I tried immunoprecipitations with Dynabeads (M-280 sheep anti-rabbit
IgG coated) and anti-p38 antibody (sc-7149), but it does not work very
well. Most of the p38 protein stays in the lysate.
thank you for the p38 kinase-assay protocol!
One further question: Which kind of sepharose beads do you use for antibody
Thank you all for help.
I wish you a happy week-end and I hope I will hear from you (perhaps) monday.