Hi, This is a general question on p-smad (2/3) detection by western. I am
trying to verify the effect of different conditions on smad phosphorylation
levels in fibroblasts (synoviocyte-like). I have tried by western blot with 2
different antibodies with tgfB induction as a positive control. I have bands
(good), but there is no significant difference in the presence or absence of
tgfB (strange - Smad phosphorylation should be increased in my cell type). I
have tried different times from 15 minutes to 90 minutes after addition of
tgfB. Compared to the litterature, I seem to have higher levels of p-smad in
the normal condition, even after starving the cells for 24H. The bands seem to
be at the expected MW. Any help would be greatly appreciated...
