RE; Isotype controls

steven micko steven_micko at email.eushc.ORG
Mon Mar 25 08:34:38 EST 1996


>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



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