RE: cytoplasmic staining

From: Calman Prussin (CPRUSSIN@niaid.nih.gov)
Date: Thu May 18 2000 - 15:39:32 EST


Matt,

Lewis Picker had a paper a couple of years ago using PFA/saponin fix and
perm and noted an IL-6 mAb was cross reacting with a histone in the cells.
They then did intracellular staining with a mAb that was generated against
the specific histone and got similar results. That would suggest that yes,
these techniques do permeabilize the nuclear membrane. Sorry.

Calman
> ----------
> From:		Matthew Morrow
> Sent:		Wednesday, May 17, 2000 11:21 AM
> To:	Cytometry Mailing List
> Subject:	cytoplasmic staining
>
>
> Flow Gurus...
>
>	Just a quick question regarding cytoplasmic staining: Do
> commercial preps such as BD's Cytofix/Cytoperm and Caltag's Fix and
> Perm also permiabilize the nuclear membrane? I'm interested in
> staining for a protein typically located within the nucleus which
> relocates to the cytoplasm when its cell is infected with an
> intra-cellular pathogen, so I need the nuclear membrane to remain
> intact to prevent false-positive signals. Any suggestions appreciated.
>
> Matt
> Matthew Morrow, MS, MT (ASCP)
> Warren Magnuson Clinical Center
> National Institutes of Health
> Bethesda, MD
> mmorrow@nih.gov/mmorrow@mail.cc.nih.gov
> (301) 496-4879
>



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