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
