[Cell-biology] Verifying levels of p-smad

Annie Lauzier via cellbiol%40net.bio.net (by Annie.Lauzier from USherbrooke.ca)
Wed Aug 20 12:26:31 EST 2008


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...



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