Isotype Controls

Scott I. Simon ssimon at bcm.tmc.edu
Wed Mar 27 11:43:57 EST 1996


Dear isotypers;
The ideal control to determine the minimum fluorescence which determines 
that an antibodie is binding specifically to its antigen is to use the 
antibody in its unlabeled form as a block. We determine nonspecific 
binding as the amount of fluorescence of the direclty conjugated mAb when 
applied to cells preblocked (to equilibrium at say 5-10 times the Kd) 
with an 'excess' of unconjugated mAb. Then you know all the 
characteristics of the mAb in terms of F/P isotype etc are the same, with 
one exception it is excluded from its antigenic site on the cell. Does 
this make sense?
Scott Simon.



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