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.ukHi, 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
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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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