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
have?

Can anyone explain this to me?

Thanks,

-----------------------------------------------------------------

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
To: histonet@pathology.swmed.edu
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??
Thanks,
Greg
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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."