[Histonet] avian/biotin block

Connolly, Brett M brett_connolly <@t> merck.com
Thu Feb 3 13:53:44 CST 2005


I've switched to the polymer-based secondaries (DAKO Envision, Vector
Impress, etc.)....biotin-free systems.


Brett M. Connolly, Ph.D.
Merck & Co., Inc.
MRL, Imaging Research
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-652-2075
e-mail. brett_connolly <@t> merck.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Emerson,
Sent: Thursday, February 03, 2005 2:32 PM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] avian/biotin block

Let me try to sum up a long story and get to my question......
Basically I am staining mouse embryos-frozen sections, for megakaryocytes.
I had trouble with endogenous AP and 15% acetic acid seemed to help. I am
having trouble with endogenous biotin, as well.  I tried Vector's
Avidin/Biotin Blocking Kit and lost my signal all together.  I am looking
for a surface antigen and I am afraid the kit blocked it all.

I would really appreciate any suggestions on blocking endogenous biotin on
frozen sections?

Also, I tried to us an AP secondary, but my signal was dimished.

Rachael L. Emerson
Center for Human Genetics and Molecular Pediatric Diseases
University of Rochester Medical Center
575 Elmwood Avenue MRBX 1.11301
Rochester, NY 14642

Tel (585) 275-5073
Fax (585) 276-0232

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