Non-saturating polyclonal antibody?

From: Art Roberts (robertar@UMDNJ.EDU)
Date: Wed Dec 29 1999 - 17:17:32 EST


Dear Flow-ers,

I am using a polyclonal antibody (unpurified immune serum) for indirect
staining which gives me a very bright signal on flow.  The problem is that
if I stain using serial dilutions of the serum (1:3, 1:10, etc.) the
signal begins to decrease with the first dilution, gradually tapers off,
and reaches the level of the negative control at about 1:10,000.  Because
the signal drops with the first dilution, I assume I have not achieved
saturation with the antibody, even though I would expect there to be a
high antibody concentration in the serum.  Does anyone have any idea
what's going on?  Thanks in advance for any help.

Art

__________________________________
Arthur Roberts
Dept. of Medicine
Robert Wood Johnson Medical School

email:  robertar@umdnj.edu
phone:  732-235-7790
Fax:    732-235-7792



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