IHC in frozen mouse liver

From:John E Morales

I am trying to establish a protocol for immunohistochemistry in 
frozen mouse liver sections in our lab.  I would first like to know 
what antibody to use as a good positive control (for liver) to 
determine whether a procedure is working in current and/or future 
experiments. I would also like to ask anyone who has successfully 
performed such an experiment to email any protocols, sources of 
information or knowledge they may have on the subject.  I'll name my 
first born after you or give you a kidney.  
    Additionally, I have tried the Vector ABC detection kit only to 
recieve a high background (even after A/B blocking) and I have so far 
not had any trouble with endogenous peroxidases showing background(is 
this normal?).  I have recently tried IHC using a SRB1 primary antibody 
and detection with an HRP labelled secondary with the results as 
follows:  
SRB1 (IgG from rabbit)= good signal
Neg.control(No primary)= no signal, very clean 
Neg.control(serum IgG from a non-innoculated rabbit)= low signal 
Seeing as my non-innoculate negative control was not clean I am unsure 
about the results.  Suggestions?  Comments?  Thank you very much for 
any help and time you provide,

JohnM.
Research Assistant II
UTatHSC





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