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 <turke...@gmail.com> Subject: [Histonet] Background in FFPE glandular tissue sections during IHC To: _histonet@lists.utsouthwestern.edu_ (mailto:histonet@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet