Re: RE; Isotype controls

steven micko (steven_micko@email.eushc.ORG)
Mon, 25 Mar 96 08:34:38 EST

>If you put two different isotypes labeled with the same fluorochrome
>in the same tube and got some nonspecific binding, then to which
>isotype would you attribute the NSB signal?

But, provided the stained cells are clearly brighter than the
nsb, does it really matter which isotype you attribute the signal to?
(ie/ In other words, can one not say of the "combined" isotype ctl
tube: This would be the MAXIMUM nsb and everything above that is
positive).

Diane,

Yes, you are right, if you include all isotypes in every tube.
Imagine, though, running a tube with an isotype antibody that did not
give much NSB. You would be assigning NSB signal from another isotype
to that antibody, and therefore getting less true postive signal from
that tube. The positive signal from one isotype antibody should not be
made to suffer for the NSB of another isotype.

Steve


Home Page Table of Contents Sponsors Web Sites
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