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I only have experience using GFP (eGFP from Clonentech), and for us the
standard 2% PFA, 0.05% Saponin didn't destroy our GFP Staining, and GFP
doesn't leak out of the cells. Haven't tried intra cellular staining on the
same time, could be tricky of course because GFP = green fluorescence, so
you would have to use PE or a conjugate for the intracellular staining..
although this works in some cases, background could be higher than with
FITC...
Sorry that I can't help you more, but maybe others have more experience in
this...
Cheers Matthias
_____________________________________________________________________________
Matthias Haury Flowcytometry Dept Immunology Institut Pasteur
mhaury@pasteur.fr Tel: 33 (01) 40 61 31 29 Fax: 33 (01) 45 68 86 39
_____________________________________________________________________________
At 15:00 -0400 2/06/97, Mark A. Miller wrote:
>Does anyone have any experience with a reporter gene, such as LacZ + FDG
>or GFP in conjunction with cytoplasmic immunostaining? My colleague
>would like to track a transient transfection in fibroblasts and measure
>elastin levels with a fluorescent MoAb at the same time.
>
>My question is this: will permeablization (to get the MoAb in) allow
>the GFP or beta galactosidase/FDG leak out? What about maintaining
>the structure/function of the reporter system? Is there a way to do
>both kinds of staining at the same time?
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
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