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GFP and Flow Cytometry

'Mike' Michael J. Moser moser at U.WASHINGTON.EDU
Thu Apr 24 13:08:29 EST 1997


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

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