IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

(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.

* Alan C. Christensen, Ph.D.          "...there are those rare  *
* School of Biological Sciences        characters who study the *
* University of Nebraska               unknown product of an    *
* Lincoln, NE 68588-0118               unknown gene.  These we  *
* Phone 402-472-0681                   call lunatics."          *
* FAX 402-472-2083                            -Sidney Brenner   *

More information about the Dros mailing list

Send comments to us at biosci-help [At] net.bio.net