[Histonet] fixation question for IHC

Anatoli Gleiberman AGleiberman <@t> cbiolabs.com
Fri Jan 28 12:33:43 CST 2011

Depends on your material, further processing and antigens. Surprisingly, some antigens are better preserved after formaldehyde fixation than after buffered formalin (probably, due to high sensitivity of particular epitope to alcohol traces from standard formalin solution). I know at least one - mouse EpCAM epithelial marker almost negative after formalin and quite well stained after formaldehyde (rat monoclonal anti EpCAM antibody G8.8 from Developmental Studies Hybridoma Bank). Morphology and antigen staining on cryo-sections from formaldehyde-fixed mouse embryos (e11-e16) are slightly better than after formalin fixation (used the same 4-6h fixation time either with buffered formalin or with 4% formaldehyde on PBS prepared from 20% formaldehyde water solution from Electron Microscopy Sciences, following PBS wash and storage in PBS at +4 C). It seems, embryonic tissue is quite sensitive to coagulation induced by the presence of small amount of alcohol. 

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 Jan Berry
Sent: Friday, January 28, 2011 1:15 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] fixation question for IHC

Is there a difference between using paraformaldehyde and neutral buffered formalin when choosing a fixative for IHC?  I would prefer to use formalin because of easier preparation, but am willing to put in the extra time to make fresh paraformaldehyde solution if there is a compelling reason.

Jan Berry
University of Michigan
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