[Histonet] Background in FFPE glandular tissue sections

[JMyers1 <@t> aol.com]

Mesruh:
The easiest method to address such is a problem is to apply a serum-free  
(e.g. casein-based) protein blocking agent, which can be obtained from a  
variety of vendors. This technique is particularly effective when the  reagent 
is allowed to incubate with the specimen for 5 to 10 minutes at RT, and  
then, rather than rinsing it off the slide with buffer, it is drained  off or 
blown off; that way, when the primary antibody solution is applied  
(immediately after this removal step), the primary has to 'work through'  the 
residual blocking agent, resulting in the binding of only  high-avidity/affinity 
antibodies to the target antigen. 
Good Luck,
Joe Myers, M.S., CT(ASCP)QIHC
 
------------------------------

Message: 3
Date: Thu, 29 Sep 2011  13:10:58 -0400
From: mesruh turkekul <turkekul <@t> gmail.com>
Subject:  [Histonet] Background in FFPE glandular tissue sections
during  IHC
To: _histonet <@t> lists.utsouthwestern.edu_ 
(mailto:histonet <@t> lists.utsouthwestern.edu) 

Dear Histonetters,

When I do IHC on FFPE tissue sections. Most of  time there is background due
to trapping of IgG/detection reagents probably  in the connective tissue
fibers, mucins or other secretions of the glandular  tissue. Especially
prostate and mammary gland. Any tips to get rid of that  kind of background?
Do you know any treatment  to block mucins or  connective tissue fibers from
nonspecifically and electrostatically  attracting IgGs and detection
reagents?

Regards,
Mesruh  Turkekul
mskcc.org