Immunohisto. question

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From:Marg Stalker <MStalker@lsd.uoguelph.ca>
To:histonet@pathology.swmed.edu
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Question regarding immunohistochemistry:  Our antigen of interest is a
protein multimer of 24 subunits, and the rabbit polyclonal antibody was
made against the subunit, i.e. the reduced form of the protein. The
problem is that the polyclonal antibody binds the intact native protein
weakly, which leads to poor immunostaining. Is there a way of reducing
the tissue proteins in situ? Reducing proteins in general requires heat
(>95 C) and a reducing agent (dithiothreitol). Any suggestions? Antigen
retrieval with trypsin is not effective.  

Thanks, 
Marg Stalker for Andy Brooks





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