RE; Isotype controls
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
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.
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