[Histonet] Re: ICC on previously Wright stained smears
TJohnson <@t> gnf.org
Mon Sep 9 12:12:48 CDT 2013
If you are getting background with little cellular staining, then the antigen retrieval method isn't working. Rene is correct that the smears you are using were most likely methanol fixed. Although heat induced epitope retrieval is thought to be most useful for formalin fixed specimens, there is also some benefit to using heat to denature (reconfigure) some of the proteins and you may get good staining in minimally fixed or alternatively fixed material. I have had success using it in minimally fixed frozen sections, when standard dogma says it shouldn't be necessary (no cross-linking occurred).
Your biggest problem may be that the antibody you are using does not recognize an alcohol-fixed protein, and no amount of antigen retrieval can fix that.
Genomics Institute for
San Diego, CA
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