Antibody titering for flow cytometry

Richard R. Hardy hardy at mighty.fccc.edu
Mon Oct 10 21:37:17 EST 1994


In article <378e40$fjg at nntp.Stanford.EDU>, ladasky at leland.Stanford.EDU
(John Ladasky) wrote:

>  I like your (Richard C. Harmon <cytact at netcom.com>) idea about using 
> anti-Fc receptor antibodies to block
> non-specific staining, but I'm not sure whether it's necessary.  Do mouse
> antibodies cross-react with human Fc receptors?  I don't expect the monkey
> cells to express Fc receptors, but I'm not so sure about Daudi.
> 

Yes, aggregated mouse Ig will light up human Ig FcR+ cells.  A simple way
to reduce this problem (at least it works for us using mouse target cells)
is to use an AirFuge to deaggregate the reagents just prior to dilution. 
We consider this standard good practice for flow staining...  Good luck.

-- 
R. Hardy
Member, Institute for Cancer Research,
Fox Chase Cancer Center, Philadelphia, PA
(215) 728-2463



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