Re: IH on Liver Tissue

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From:Jeff Crews <jcrews@organo.com>
To:kkdulany@unmc.edu, histonet@pathology.swmed.edu
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     With extremely bloody tissue like spleen or liver, it is sometimes 
     impossible to quench all of the peroxidase present in the blood. Try 
     an alkaline-phoshatase detection system.
     
     Jeffrey Crews, HTL (ASCP)
     Organogenesis, Inc.


______________________________ Reply Separator _________________________________
Subject: IH on Liver Tissue 
Author:  <kkdulany@unmc.edu> at internet
Date:    07/10/2000 11:03 AM


How can you successfully perform immunohistochemistry on slides of liver 
tissue using horse-radish peroxidase and DAB?  How do you quench the 
endogenous peroxidase in the tissue so everything doesn't turn brown?  I've 
tried 3, 10 and 15 % H2O2 in both methanol and di water for 20 min. up to 1 
hour with no success.  The tissues are formalin fixed and embeded in 
paraffin.  We then used a pH 6.0 citrate buffer/microwave antigen retrieval 
system for our primary Ab detection.  Even with all that the endogenous 
enzyme caused the false positive with the DAB.  Anyone have any 
suggestions?
Thank you for all replies.
Karen in Omaha
     
     
     




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