Need help with C12FDG (Imagene green) for lacZ

rbram at cris.com rbram at cris.com
Sun Nov 19 00:55:53 EST 1995


Has anyone gotten this to work?? I'm trying to detect lacZ expression
in Jurkat T cells using this derivative of FDG (fluorescein di-galactoside)
from Molecular Probes. I can get regular old FDG to work, of course, so
I know my cells are expressing lacZ. But there is no fluorescence at all
when I try the "new and improved" Imagene Green.
I've tried adding it to cells at 37 degrees at 50 microM and also at 100 microM.
(for one hour.)

The point of the "improved" reagent is that it is supposed to be retained
within the cell once it is cleaved and becomes fluorescent, unlike FDG which
is impermeable until cleaved and then only stays in cells at zero degrees.



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