Re: Thanks re:endogenous peroxidase, a serum blocking question

<< Previous Message | Next Message >>
From:Tim Morken <timcdc@hotmail.com>
To:HistoNet@Pathology.swmed.edu
Reply-To:
Date:Tue, 06 Apr 1999 11:38:26 -0400 (EDT)
Content-Type:text/plain

Bruce,

We use a 20% sheep serum (in tris-saline-tween20 buffer) before the 
primary. We dilute the primary in the same serum-buffer, so it 'keeps on 
blocking.' We don't rinse the blocking serum before putting on the 
primary, only an air-blow (using the DAKO auto-stainer). We then rinse 
the with the tris-saline buffer without the serum.

If you have a lot of background problems then a second serum blocker may 
be necessary before applying the secondary antibody. In general, I would 
rather keep the procedure simple and use as few steps as possible.

Tim Morken, B.A., EMT(MSA), HTL(ASCP) 
Infectious Disease Pathology
Centers for Disease Control
MS-G32
1600 Clifton Rd.
Atlanta, GA 30333
USA

email: tim9@cdc.gov
       timcdc@hotmail.com

FAX:  (404)639-3043




----Original Message Follows----
From: Bruce A Rasmussen <brasmuss@osf1.gmu.edu>
To: HistoNet@Pathology.swmed.edu
Subject: Thanks re:endogenous peroxidase, a serum blocking question
Date: Tue, 06 Apr 1999 10:44:30 -0400 (EDT)

My thanks to all who wrote back with detailed responses to my endogenous
peroxidase question. If I could trouble folk with just one more 
question. 
I have a protocol that calls for a 5% serum block (30 min to the 
secondary
animal) before and primary and before the secondary. There is much 
debate
about whether the second block is necessay, if it will reduce the 
signal,
or if rinsing after the serum blocks is good or bad. Some folk also
suggest that a couple of percent BSA is a good idea.
Thanks again,
Bruce 



--------------------------------------
Bruce Rasmussen
Predoc Fellow in Experimental Neuropsychology
The Krasnow Institute for Advanced Study
George Mason University
Mail Stop 2A1
Fairfax, VA 22030-4444
Office:703-993-4358
Lab:703-993-4369
Home:703-765-4570
Fax:703-993-4325
---------------------------------------





Get Your Private, Free Email at http://www.hotmail.com



<< Previous Message | Next Message >>