Nick,
Fix the brains in Pharmingen Zinc Tris, process to paraffin and use the
Pharmingen antibody. Works like a charm.

Brett

Brett M. Connolly, Ph.D.
Merck & Co., Inc.
MRL, Imaging Research
WP-44K
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-993-6803
e-mail. brett_connolly@merck.com


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon D.
Bucana
Sent: Wednesday, June 01, 2005 9:44 AM
To: Donin, Nick (NIH/NCI)
Cc: 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] mouse endothelial cells in brain


The only time we got good staining is on frozen sections using anti-mouse 
CD31 from either Pharmingen or Serotec in both immunofluorescence and DAB 
or AEC staining.


At 11:54 AM 5/31/2005 -0400, you wrote:
>Histonetters,
>
>This is my first posting, so I'm excited that someone out there might be
>able to help me.
>
>
>I am trying to stain endothelial cells in brain tumors in formalin fixed
>paraffin embedded mouse tissues.  I have tried several antibodies, and none
>of them seem to work.  I am consistently getting high background and no
true
>staining.  I have used the following antibodies:
>
>
>
>BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355
>
>Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this
>works, but my protocol didn't produce good results)
>
>Cedarlane mouse CD31 cat#CL8930 AP
>
>Cymbus Biotechnology anti-ms CD31 cat# CBL1337
>
>
>
>The protocol I use is shown below.
>
>
>
>
>
>I'm confident that out of these antibodies, at least one will be able to
>stain the endothelial cells in these tumors, but I think that my protocol
>may be the problem.  Could someone possibly suggest and antibody or
protocol
>that would work for me?  Thanks so much for your help, much appreciated.
>
>
>
>Nick Donin
>
>CRTA
>
>Neuro-Oncology Branch
>
>National Cancer Institute
>
>National Institutes of Health
>
>9000 Rockville Pike
>
>Building 35, Room 2B-203
>
>Bethesda, MD 20892
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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