[Histonet] Albumin and Epcam to stain mouse cell pellets

Eva Permaul eca9 <@t> georgetown.edu
Tue Sep 25 13:52:48 CDT 2012

Thank you Anatoli.

Let me add some more info to my question. The samples are FFPE.

On Tue, Sep 25, 2012 at 2:43 PM, Anatoli Gleiberman <
AGleiberman <@t> 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: 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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