[Histonet] BRDU

[Andrea Marion]

Yes, I am also using pH 6 HIER with 10mM sodium citrate, 0.05% Tween-20. I
do 20 minutes at 95 degree C to retrieve BrdU on mouse embryo tissues. I
use Abcam ab6326. Their product sheet states that you may used heat for
antigen retrieval.

The BrdU epitope needs to be revealed by denaturing the double-stranded
DNA - and you can do that with an enzyme, acid, or heat - so if you are
already doing HIER, the acid/enzyme treatment can be unnecessary (and may
even damage the tissue/other antigens you are interested in).

Andrea Marion
Graduate Student
University of Illinois at Chicago



I used pH 6 HEIR instead of HCl/enzyme for BrdU - it works very well on
mouse and monkey intestine. Did not switch to HCl after that. But now I
prefer to use EdU instead of BrdU (Click-iT kit from Invitrogen). It is
more expensive, but much more sensitive, fast, easy to use and well
compatible with all range of antigens(for multi-color fluorescence).
Unfortunately, there are no enzymatic reaction for EdU - only
fluorescence.

Anatoli Gleiberman, PhD
Director of Histopathology
Cleveland Biolabs, Inc
73 High Street
Buffalo, NY 14203
phone:716-849-6810 ext.354
fax:716-849-6817
e-mail: AGleiberman <@t> cbiolabs.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C B
Sent: Wednesday, June 08, 2011 8:37 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] BRDU

Is anyone using ph6 HEIR instead of HCl/enzyme digestion for BRDU staining?
I've read 2 manufacturer datasheets that recommend pH 6 HEIR.  I thought the
routine pretreatment was HCl/enzyme digestion.  We have FFPE rat intestine to
stain. Any feeback is appreciated.
Cindy Baranowski, HT
SJTRI
Atlanta, GA
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