RGFP and FACS in mammalian cells

Dezz Ropp dezz at gene.com
Wed Nov 29 12:33:29 EST 1995

------------ Original Message -------------
Clontech showed a beautiful picture of FACS on cells transiently
tranfected with the new red shifted GFP clone. Does anyone have
'homegrown' results with flow cytometric analysis?

I am planning to use GFP as a reporter in a cAMP inducible promoter=20
construct and assay by FACS. Doe anyone have experience with something=20
remotely similar? Thanks.


Dr. David de Graaf
Department of Membrane Research and Biophysics
Weizmann Intitute of Science
Rehovot, 76100
tel * 972 8 343 686
fax * 972 8 344 112

--------- End of Original Message ---------

------------ Original Message -------------
I am also planning on sorting cells on FACS following expression in
cells (Rat-1 cells).  In my case I will be using the GFP on a CMV
promoter to
use as a marker for cells successfully co-transfected with
kinase constructs.  I plan to use the FACS-sorted cells for MAP kinase
I, too, would be interested whether anyone has information regarding
approach, especially the conditions under which expression of GFP would
adequate for its use as a FACS-sorting marker.  Thanks,


Dr. Bruce Magun
Department of Cell and Developmental Biology
Oregon Health Sciences University
Portland, OR 97201
tel 503-494-7811
fax 503-494-4253
--------- End of Original Message ---------

We have been successful in FACS analyzing both wt GFP and S65TGFP in=20
various mammalian cell types. For a detailed description please see=20
our manuscript in the upcoming issue of Cytometry: Ropp, J. D., et al.=20
(1995). =D2Aequorea green fluorescent protein (GFP) analysis by flow=20
cytometry.=D3 Cytometry 21(4). I hope this can provide some answers.


Dr. Dezz Ropp
Genentech, Inc.
460 Pt. San Bruno Blvd
S. San Francisco, CA 94080
Fax: 415.225.5335

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