Re: Isotype Controls
Scott I. Simon (ssimon@bcm.tmc.edu)
Wed, 27 Mar 1996 10:43:57 -0600 (CST)
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.
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
