Hi Tim,
I'm with you, that's exactly what I thought as well. But these guys 
were working on formalin-fixed, paraffin-embedded tissues!?

So I'm thinking the permeabilizing step is not necessary. I'm going 
to try it without any treatment and perhaps with a heat retreival 
method and compare results. Thanks for the reply.
Greg

Date sent:      	Tue, 06 Aug 2002 13:41:45 -0400
From:           	"Morken, Tim" 
Subject:        	RE: Ag retreival vs. Permeabilizing
To:             	'Greg Dobbin' , histonet@pathology.swmed.edu

> Detergents such as Triton and saponin "permeabilize" cell membranes by
> making holes in them. This is a physical process that simply makes a "door"
> for the antibody to enter the cell. Usually this is done on whole-cell preps
> such as cell smears and cytospins in which the cell membrane is usually
> intact. It can be done on frozen sections, however it is usually not done on
> paraffin-embedded tissue since that is normally fixed. The process is very
> different than the "antigen retreival" techniques we use on paraffin
> sections which are designed to break formalin/protein bonds. 
> 
> Tim Morken
> CDC, Atlanta
> 
> 
> -----Original Message-----
> From: Greg Dobbin [mailto:dobbin@Upei.CA]
> Sent: Tuesday, August 06, 2002 12:55 PM
> 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."


~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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."