Hi all;

Rat anti-mouse CD31 (MEC13.3) from Pharmingen works well in FFPE mouse
tissues.  Enzyme digestion with Trypsin - overnight primary antibody
incubation at 4C- secondary rabbit anti-rat IgG antibody 30 mins at RT-
Vectastain Elite ABC 30 minutes  RT -NEN biotinylated tyramide amplification 3
mins RT - followed by another incubation in Vectastain Elite ABC 30 minutes at
RT.  Make sure you also block endogenous biotin with Avidin/Biotin Blocking
Kit, endogenous peroxidases with H2O2 and endogenous proteins with normal
serum.

Barb Wright
Genentech, Inc.

"L. Muffley" wrote:

> This article has been very helpful for me:
> "Adenoviral Expression of Vascular Endothelial Growth Factor -C Induces
> Lymphangiogenesis in the Skin" Enholm et al 2001
> (Circ Res. 2001; 88:623-629)
>
> I am currently troubleshooting staining my FFPE mouse skin with RDI's
> PECAM/CD31 rat anti-mouse monoclonal.
> The paper above used tyramide amplification to enhance staining
> (they used Pharmingens PECAM-1).
> Also-When I contacted the researchers in Finland they said that they used
> trypsin digestion (45min) - and no HIER.
> So I'm testing with those perameters currently.
>
> And to quote Gayle Callis from a previous post
> "Murine CD markers often hate NBF much to mouse people's dismay and
> frozens are the only recourse"
> True, so true...
> Our lab has great success with PECAM in Zinc (tris) fixed mouse skin.
> But alas, I have an entire FFPE mouse experiment that I need to stain with
> an endothelial cell marker.
> I'm hoping for success with trypsin and TSA.
>
>                                 Lara Muffley
>                                 Dermatology Dept
>                                 University of Washington
>                                 Seattle, WA
>                                 muffley@u.washington.edu
>
> On Tue, 23 Oct 2001, Edwards, R.E. wrote:
>
> >
> >
> > --
> > I  am  looking  for  an  antibody  to  mouse  endothelial  cells  that
> > works  on  paraffin  processed  tissues, any  suggestions  gratefully
> > received.
> >                                 Thanks
> >                                        Richard  Edwards
> >                                                    MRC TOXICOLOGY  UNIT
> >                                                             LEICESTER
> > U.K...........
> >
> >