Non-green fluorescent protein

Andrew Doherty A.Doherty at Bris.ac.uk
Thu Nov 28 03:27:59 EST 1996


Gene Regulation wrote:
> 
> I would like to use a fluorescent protein as a marker for DNA uptake in
> transient assay (mammalian cells) but need to counterstain the cells for
> a second protein with FITC ie green! Is there a red or blue fluorescent
> protein which is easy to see using a fluorescent microscope?
> 
> Secondly, I have tried using S65T with little success in COS and 3T3
> fibroblasts. Fluorescence is there but is weak - we transfect with
> sv40-driven S65T and 24 - 48 hours later look at the cells (growing on
> slides) under FITC illumination - the cells are still in DMEM and are
> viable. Can anyone give me a good protocol for using GFP?
> 
> Many thanks
> G Atherton
> reply to grggta at picr.cr.man.ac.uk
Hi, we have no problem in getting GFP fluorescence in either HEK293 or
CHO cells with the S65T mutant, but using a CMV driven vector and
standard CaPO4 precipitation. Do other genes transfect in your cell
lines?

Andy
-- 
*************************************************************
Dr Andrew Doherty		email -  a.doherty at bris.ac.uk
Dept. Anatomy			Tel (0117)9287421
School of Medical Sciences	Fax (0117)9287402
University of Bristol
University Walk
Bristol UK
BS8 1TD
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