Problem with nonspecific antibody binding

Dennis Goode goode at ZOOL.UMD.EDU
Wed Feb 26 11:49:45 EST 1997


Homer,

Sometimes sites exist that bind antibodies non-specifically.
Try blocking with unlabeled  antibodies or  serum from the same 
species as your secondary antibody e.g. 2% non-specific goat serum (in addition to 
BSA).
Tween 20 sometimes helps at about 0.1% in the blocking 
solution, but also might remove some antigens.

-Dennis

To:            cellbiol at net.bio.net
From:          divernlb at aol.com (DiverNLB)
Subject:       Problem with nonspecific antibody binding
Date:          26 Feb 1997 00:31:15 GMT

Hello. I am currently developing a protocol for immunofluorescent
detection of the stress proteins hsp70 and cpn60 on cryosectioned Mytilus
mantle tissue. The problem I am having is nonspecific binding of the
secondary antibody (conjugated to fluorescein), leading to strong
false-positive signals. I have tried blocking with bovine serum albumin,
fetal bovine serum, and horse serum, with little or no effect. Could
anyone suggest any other solutions? Please respond to DiverNLB at aol.com.
Thank you.


Homer O. Ellison
Dept. of Biological Sciences
California State University, Long Beach


Dr. M. Dennis Goode                 Phone (301) 405-6917
Department of Zoology               Fax   (301) 314-9358
University of Maryland              e-mail goode at zool.umd.edu
College Park MD 20742
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"If the Lord Almighty had consulted me before embarking upon the 
creation, I should have recommended something simpler."
            -Alphonso X of Castile, 15th Century



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