RE: Ag retreival vs. Permeabilizing - extra question.

From:Phil Bergin

We use saponin (or triton) to permeabilize cells for intracellular
staining of cytokines in the cell cytoplasm etc.

According to the people who developed our lab protocol it does make a
difference, but the question always asked is why is it necessary? We cut
cryosections at 8 microns - does this 'cut through' cells exposing the
inside of some cells.  If is does then what effect does the saponin

Can anyone explain this to me?



Philip Bergin
Göteborg University
Department of Medical Microbiology and Immunology
Box 435, SE-405 30 Göteborg, Sweden
Phone: +46-31-3424727
Fax: +46-31-826976

-----Original Message-----
From: Greg Dobbin [mailto:dobbin@Upei.CA] 
Sent: den 6 augusti 2002 18:55
Subject: Ag retreival vs. Permeabilizing

Hello All,

I found a "Brief Communication" in Vet Pathology (38:116-119; 
2001) that discusses expression of COX-2 in canine renal 
carcinomas. The authors do not mention antigen retreival, rather, 
they speak of permeabilizing the tissues using triton and 
saponin. Is permeabilizing (as described) a form of antigen 
retreival or does it serve another purpose?

Has anyone else heard of this before? Explanation??
Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 University Ave.
Charlottetown, P.E.I.
Canada,  C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851
"A farmer is a person outstanding in their field."

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