(no subject)

Alan C. Christensen, Ph.D. achristensen at unl.edu
Mon Sep 11 12:12:25 EST 1995


Here's a simple question, which I hope someone has a simple answer to.  
We are staining whole-mount embryos with an antibody, and then with an 
HRP-conjugated second antibody.  We are getting signal, but there is 
some background, presumably from endogenous peroxidases.  The standard 
protocol to inactivate peroxidases is to incubate with 3% H2O2 in 
methanol, just after devitellinizing.  However, the antigen is methanol 
sensitive, so we are hand-peeling the embryos.  I've been told that 
treatment of specimens with 3% H2O2 in aqueous solutions results in 
violent bubbling and destruction of the specimen.  We will be trying a 
few things, but if anyone has a successful method for inactivating 
endogenous peroxidase without methanol, please let me know.

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* Alan C. Christensen, Ph.D.          "...there are those rare  *
* School of Biological Sciences        characters who study the *
* University of Nebraska               unknown product of an    *
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