I have some problems with the western blot when I want to probe other antibody. When I make the western for my protein there aren't problems and I obtained a really strong signal, but when we reproble the membrane for the control antibody there isn't any signal.
My stripping protocol is:
Tris-HCl 0.5M pH 6.8
I put my membrane with this buffer during 30 min at 60 ºC.
Then I wash with PBS-tween during 1 h changing the buffer.
I reblocked the membrane with PBS-tween-milk and incubate with the primary antibody.
This protocol worked in the past and I don't know where the problem is now.