[Histonet] Albumin and Epcam to stain mouse cell pellets

Anatoli Gleiberman AGleiberman <@t> cbiolabs.com
Tue Sep 25 13:43:14 CDT 2012

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
e-mail: AGleiberman <@t> cbiolabs.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> 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
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