Re: BrdU

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From:Katie B <> (by way of histonet)
To:histonet <>
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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!


Catherine "Katie" Bresee Bennett
Laboratory for Experimental Pathology
Department of Veterinary Pathology
Michigan State University

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