Young-Joon KIM,
Perhaps the most difficult aspect of affinity methodologies is the
non-specific background. One approach to eliminate this is to "pre-clear"
the lysate prior to adding the photoaffinity reagent. Pass the lysate
through a streptavidin column first, then add your reagent. thia should
eliminate many of the interfering bands on your western.
----------------------------------------------------------------------------
-------
Gordon Alton, Ph.D.
Lead Scientist
Assay Development and Analytical Protein Chemistry
Department of Informatics and Functional Genomics
Signal Research Division of Celgene
5555 Oberlin Drive
San Diego, CA 92121
Email: galton@signalpharm.com
Phone: 858-558-7500 x8252
Fax: 858-623-0870
WWW: http://www.signalpharm.com
----------------------------------------------------------------
-----Original Message-----
From: Young-Joon KIM [mailto:yjkim@ucrac1.ucr.edu]
Sent: Sunday, February 04, 2001 4:44 PM
To: Recipients of ABRF List
Subject: Anti-Biotin Antibody
Hello~:
I am trying phtoaffinity labeling with using biotinylated photo-activatable
crosslinker. Recently I found that many protein from tissue (CNS) can bind
with stretavidin-HRP even without being labeled with cross-linker. So I want
to use anti-biotin antibody instead of avidin/streptavidin to detect
biotinylated protein in western blotting. Could you recommend the
commercially available good anti-biotin antibody (in terms of specificity)?
I found some of them in catalogues of Pierce and Jackson Lab.
Many thanks,
Young-Joon KIM
UC Riverside
This archive was generated by hypermail 2b29 : Fri Feb 23 2001 - 13:03:34 EST