Quantitative flow in the nucleus

From: Lars Brichta (LBrichta@gmx.net)
Date: Fri Nov 15 2002 - 04:33:58 EST


Hi flow-ers,

I've got a question referring to quantitative flow cytometry. I wanna
measure an antigen in the nucleus and can't decide between FITC and PE for antibody
labeling (I wanna use a directly labeled primary antibody). I know there is
a lot you can't consider in advance, so as antigen access, impact of fixation
and permeabilization methods.....but still -  I think PE seems to be more
sensitive because of the number of fluorophores each molecule shows, but at the
same time the molecule is much bigger, FITC seems to be less sensitive but
it is smaller and might have less negative influence on the access to the
antigen.
What do you think?

And, another thing: Does anybody know a good nuclear protein standard I can
use? I intended to use histone proteins, but could not find a monoclonal
antibody against any of the histones....


Thanks, I really appreciate your help!

Lars Brichta
University Clinics Bonn, Germany

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