I have also used the pharmingen cd31 by frozen in mouse brain, muscle, and
skin. It works extremely well in fresh system.  FFPE on the other hand is
much trickier. It works but very inconsistently, fixation and tissue
processing are the key variables.  Once you have obtained staining in FFPE,
you need to follow the same sacrificing (perfusion etc..) grossing,
processing procedure with "religious fanaticism"  (no offense to
anyone.....)

L

____________________________________
Luis Chiriboga Ph.D.
NYU Cancer Institute and
Bellevue Hospital Center
New York University School Of Medicine
Department Of Pathology 4W27
462 First Avenue
New York, N.Y. 10016
W(212) 562-4667.
F(212) 263-2041






-----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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