On a related topic, does anyone know if the Biocare rat CD31 clone works for immunofluorescence on FFPE sections using a directly conjugated secondary antibody?
Thanks, Adam On Tue, Feb 23, 2010 at 1:06 PM, Andrea T. Hooper < [email protected]> wrote: > Hi Phebe, > > In my experience that CD31 clone you refer to doesn't work well mainly b/c > of paraffin embedding - in combination with PFA/formalin fixation. On > PFA/formalin fixed frozen samples it works just fine with a trypsin antigen > retrieval step. I find it's the paraffin that is the real killer. Try > Biocare's anti-CD31, that works on FFPE. Or use another marker of > endothelium (if that's what you are using CD31 for). > > Alternatively, are you sure your second antibody only works in FFPE > formalin fixed tissue? Because it may well work on formalin fixed frozen > tissue, then you can use both on that instead (just remember to do the > trypsin digestion). Or use fresh snap frozen tissue postfixed in PFA for 10 > minutes then no antigen retrieval necessary. > > Good luck! > Andrea Hooper > > > --- Phebe Verbrugghe <[email protected]> wrote: > > > Hi all, > > I would like to do an immunofluorescent double labeling > with two > antibodies but 1 antibody works on acetone fixed frozen > tissue but not > on formalin fixed paraffin embedded tissue (CD31 BD > pharmingen 553370) > and the other one works on formalin fixed paraffin > embedded tissue but > not on acetone fixed frozen tissue. Is there any way I > could still do a > double labeling and how? > Also, does anyone have experience with zinc fixative? If > my antibody > works on formalin fixed tissue is it likely to also work > on zinc fixed > tissue? > > Thank you very much in advance, > > Phebe > > > > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
