[Histonet] Re: PECAM-1 antibody

koellingr <@t> comcast.net koellingr <@t> comcast.net
Sat Jan 26 13:53:21 CST 2008

I agree completely with Carl, in asking the question about using CD31 which is a question that must always be asked for any project as far as just following conventional wisdom.  Different needs and different data and according to your model might require different reagents. CD31 is kind of a default gold standard for endothelial cells but not perfect.  And a more difficult antibody for mice/rats.  Have used BDPharmingen rat-anti mouse CD31 on FFPE mouse tissue, WITH TYRAMIDE AMPLIFICATION, and it worked quite well.  Detailed this several years back on HistoNet.  Biotinylated rat Abs works on rats with same caveat of amplification need.  Never did like that Santa Cruz CD31, even when popular back then, for specific and general reasons.  Aquaporins in endothelial cells but also myocytes, basal layers of some epithelium, etc.  Laminins not sure what that would do? or how helpful; is an extracellular matrix in basement membrane and other places.  CD34 (mucosialin) might be great choic
e for capillary endothelial cells but is in other places (hematopoietic stem cells).  vWF might be great choice, works in FFPE nicely, but not ALL endothelial cells synthesize vWF.  I've used VE-cadherin (vascular endothelial-cadherin) in FFPE mice for endothelial cells.  In short there are some good choices out there for FFPE mouse/rat endothelium and while it might be a bit more tricky and problematic than easier and more focused targets with easier reagents, better than hoping for the reincarnation of that dumb goat that has been diefied, but whose anti-CD31 antibody containing serum I was always cautious and wary of in the first place.

Ray Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "Carl Hobbs" <carl.hobbs <@t> kcl.ac.uk> 

> Interesting. 
> Just out of interest, why do you want to use Pecam 1? 
> I have not been able to get same sc Ab to work on FFPW sections of 
> mouse/rat endothelial cells. 
> However, I understand that it is also positive for non-endothelial cells so, 
> is this the best marker for endothelial cells? 
> Any views on using anti CD34, Aquaporin, VWF on mouse/rat FFPW sections to 
> detect capillaries? 
> Can anti laminin be used instead? 
> Carl 
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