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
