primary stopped working for IHC - immunohistochemistry (Sep/30/2004 )
I've been using a primary antibody (anti-BMP4 from R&D Systems) for immunohistochemistry on mouse testis tissue. Everything was working fine, then all of a sudden, it stopped working. I've been using all the same reagents and conditions, and I havent changed my batch of antibody (same batch but different aliquots that I reprared myself when the antibody arrived). As far as I can see, nothing has changed in the protocol, yet I cant get it to work.
I was wondering whether this sort of thing is common? I dont have alot of experience with immuno work and would like to know it this is to be expected.
have you been doing a double immunostaining? The second Ab, or enzyme might have failed. If you haven't checked it yet, try IHC with different primary Ab about which you are sure that is working. I am not an expert, so my advices may be useless