Alcohol fixation will rapidly destroy GFP fluorescence. I use 1/20th
volume of formalin for 5 minutes at RT. Cells will remain fluorescent for
several days stored in PBS at 4 C. WT GFP is essentially useless. Human
codon bias optimized S65T (or other enhanced) GFP is highly recommended
for FACS w/ human cells.
Mike Moser Tel: 206-616-7391
UW Department of Pathology FAX: 206-543-3644
Box 357470 moser at u.washington.edu
Seattle, WA 98195 http://weber.u.washington.edu/~moser
On 24 Apr 1997, Simon D. Scott wrote:
> Hello. We're investigating the use of wild-type and mutant GFPs in live
> and fixed cells (e.g. MCF7 breast tumour line) and wish to quantitate
> using FLOW CYTOMETRY. We are counterstaining fixed cells with PI. 70%
> ethanol is the fixative.
> Any advice? - Simon Scott/Brian Marples PICR