I would consider using a non-biotin detection system such as Hrp Labelled
Polymer, there are several out there now, the one I use is from DAKO called
Envision.
Patsy 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emerson,
Rachael
Sent: Thursday, February 03, 2005 12:32 PM
To: 'histonet@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.
Thanks
Rachael 

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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