Re: BrdU
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From: | Katie B <bresee98@yahoo.com> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
I have a tried and true protocol for staining for BrdU in
paraffin-embedded tissues with consistent results. Having the tissues
fixed in Zinc Formalin really makes a huge difference in the quality
and consistency of the stain. The staining protocol calls for
Proteinase K and acid treatment steps to permeabelize the cells and
denature the DNA. Other than that I use anti-brdu from Becton
Dickinson, and a mouse IgG Vectastin Elite kit from Vector Labs.
I agree that Sharron's problem between the staining quality in the
intestines and the vessels is probably because the intestines simply
have more BrdU incorporated into the cells. While I don't know how
rapidly your blood vessel cells are proliferating, intestines pick up
BrdU very well. This is assuming that you are doing a Protease and an
acid pre-treatment steps in your staining. If not, adding these steps
might change the staining intensity/quality.
PS - I've brought up BrdU staining in the past for plastic-embedded
tissues and several people asked me for the protocol I was using. I
want to add a disclaimer to the protocol I sent out that I have never
actually done it myself until just this week and I'm having problems
with non-specific staining. Will keep any interested persons posted
on my quest!
-Katie
==
Catherine "Katie" Bresee Bennett
Laboratory for Experimental Pathology
Department of Veterinary Pathology
Michigan State University
*new* e-mail: bresee98@yahoo.com
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