Thank you Anatoli. Let me add some more info to my question. The samples are FFPE. Thanks, Eva
On Tue, Sep 25, 2012 at 2:43 PM, Anatoli Gleiberman < agleiber...@cbiolabs.com> wrote: > Eva > For mouse EpCAM I have used routinely during past 15 years G8.8 rat > monoclonal antibody from Developmental Studies Hybridoma Bank. Concentrated > supernatant (1:200 dilution for IF) works very well on 4% paraformaldehyde > fixed frozen samples (or on sections from fresh-frozen samples post-fixed > with formaldehyde). Epitope is very sensitive to alcohol, so even fixation > with NBF instead of formaldehyde diminishes staining dramatically. I was > told that this antibody works on formalin-fixed paraffin sections after > HIER retrieval, but never got results comparable to frozen sections. Don't > have any recommendations regarding albumin, I used home-made antibodies for > that purpose many years ago. In general, paraformaldehyde fixation and > frozen sections worked satisfactory. Classical approach for albumin and > alpha-fetoprotein staining was to fix in Saint-Mary fixative (1% acetic > acid in absolute ethanol) following routine paraffin embedding. > Nevertheless, short fixation with NBF (4h at room temp, following PBS wash) > and paraffin embedding works as well. Best results regarding liver secreted > proteins (as well as all membrane and cytoskeletal markers) could be > achieved on cryo-sections after fixation by perfusion with 4% > paraformaldehyde in PBS. > > Anatoli Gleiberman, PhD > Director of Histopathology > Cleveland Biolabs, Inc > 73 High Street > Buffalo, NY 14203 > phone:716-849-6810 ext.354 > fax:716-849-6817 > e-mail: agleiber...@cbiolabs.com > > -----Original Message----- > From: histonet-boun...@lists.utsouthwestern.edu [mailto: > histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Eva Permaul > Sent: Tuesday, September 25, 2012 1:21 PM > To: histonet > Subject: [Histonet] Albumin and Epcam to stain mouse cell pellets > > Good afternoon, > Has anyone used Albumin and Epcam antibodies to stain mouse samples? If > you have would you please share the antibody information and conditions > with me? > Thank you, > Eva Permaul > Georgetown University > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet