[Histonet] mouse endothelial cells in brain

[Donin, Nick (NIH/NCI)]

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