Antibody titering for flow cytometry

Richard R. Hardy hardy at mighty.fccc.edu
Thu Oct 6 13:00:17 EST 1994


In article <36uq0b$58m at nntp.Stanford.EDU>, ladasky at leland.Stanford.EDU
(John Ladasky) wrote:

> Greetings, everyone,
> 
>         Using a flow cytometer and an antibody at various dilutions, I've
> just analyzed the fluorescence intensity of a particular cell-antibody 
> combination.  I now have a nice, asymptotic curve.
> 
>                             Mean
>         ug MAb added    Fluorescence
>         ----------------------------
>             0.00               8
>             0.63             364
>             1.25             541
>             2.50             812
>             5.00            1050
>         
>         Now what?  At what level is an antibody considered to be "titered"?
> Any advice or references would be greatly appreciated.  Thanks!
> 

You're close to some REAL experts on Flow Cytometry and cell surface
immunofluorescence - why not visit someone in the Herzenberg lab / FACS
development group (in the basement of CMGM) and show them your plots? 
Aaron Kantor may still be around...  We could give advice and recommend
chapters/books, but it's still good to see the raw data - i.e., to
recognize potential pitfalls like problem backgrounds, etc...

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



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