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RGFP and FACS in mammalian cells

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


------------ Original Message -------------
Hi,
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.

David

Dr. David de Graaf
Department of Membrane Research and Biophysics
Weizmann Intitute of Science
Rehovot, 76100
Israel
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
mammalian
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
dominant-negative
kinase constructs.  I plan to use the FACS-sorted cells for MAP kinase
assays.
I, too, would be interested whether anyone has information regarding
this
approach, especially the conditions under which expression of GFP would
be
adequate for its use as a FACS-sorting marker.  Thanks,

Bruce

Dr. Bruce Magun
Department of Cell and Developmental Biology
Oregon Health Sciences University
Portland, OR 97201
USA
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.

dezz


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



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