Re: CD31/pecam in mouse

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From:denise M m Long-Woodward <> (by way of histonet)
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Thanks for your reply. We are getting great staining with our CD31's. The
problem is the researcher wants an antibody that is exclusive to
endothelial cells of mouse ( monocytes, etc.)  and it should not
cross react with human endothelial cells. (it's a xenograft, human on
mouse). Any ideas are appreciated.  Denise Woodward

On Fri, 3 Dec 1999 08:49:17 -0700 Gayle Callis
<> writes:
> Have a grad student who does this more than frequently, now doing it
> with confocal.  Frozen sections, cut at 5 um, cold acetone fixation
> for 10 min, refrig temps, air dry 30 min before fixation, fix then
> air dry for another 30 min.  Staining is done (you will need to do
> a dilution panel), using Dulbeccos PBS, pH 7.4 with 0.2% goat serum,
> block is 5 - 10% goat serum 30 min,  staining with antibody is 30
> min,
> Secondary is goat antiRat F(Ab')2 biotin, 1:250 from Biosource
> (Jackson
> has excellent secondary also), Strepavidin-HRP 1:500 from Biosource
> 20 min,
> then chromogen of choice, we like Vector AEC or Pierce enhanced DAB.
> Go easy on hematoxylin counterstain.  If you can add Tween 20 to
> buffer,
> 0.05% to 0.1% and we do both peroxidase block (Dako) and avidin
> biotin block)
> Good luck
> Gayle Callis

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