[Histonet] double staining

[Gayle Callis]

The shielding can be done by chromogens too, Chris van der Loos teaches this 
in his multiple staining course.  He always develops the Alk phos first, 
then the HRP.  Hopefully he is looking in on this messaging too.

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT
----- Original Message ----- 
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, July 14, 2008 4:21 PM
Subject: RE: [Histonet] double staining


Have you tried doing the two antibodies in reverse order?  Sometimes the two 
antigens are configured such that the binding of antibodies to one blocks 
the access of antibodies to the other.  Picture it as one antigen protruding 
farther from the cell surface than the other.  If you stain the "taller" 
antigen first, it can form a protective shield, so to speak, that prevents 
antibodies from penetrating to the underlying "shorter" antigen.  But, if 
you stain the "shorter" antigen first, then the "taller" one may still 
protrude far enough for its anitibody to find it.
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