Hi Mark, 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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