[Fluorescent-proteins] Quenching Fluorescent Proteins

Confused Dave via fluorpro%40net.bio.net (by confuseddave from gmail.com)
Fri Nov 21 05:55:09 EST 2008


Hello,

I'm looking for a way to reliably inactivate fluorescent proteins
(EGFP and dsRed) without harming epitopes for immuno.

A little background:  I'm electroporating inducible constructs into
the chick embryo.  We use dsRed as the electroporation control, and
EGFP as a marker for activation of our doxicyclin-inducible construct.
 We want to use immunofluorescence to on these samples.  We currently
use 4% formalin to fix, washes with PBS+tween, usually ethanol
dehydration for storage, and sucrose-agar embedding for sectioning,
and the fluorescence still comes up strong.  Obviously, imaging with
epifluorescence with a green and red channel, we can't add another
fluorophore.

Best case would be a way to specifically kill the RFP leaving the
green intact, although as long as we can recover the GFP with an
antibody, this isn't a problem.  Also looking for something that won't
damage our epitopes to badly!

Thanks,
Dave



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