(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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