Fixing EGFP-expressing cells for FACS

'Mike' Michael J. Moser moser at U.WASHINGTON.EDU
Fri Nov 7 19:06:46 EST 1997


Jacqueline

I fix cells expressing S65T GFP by addition of 1/20th volume formalin (the
cheap liquid, just replace your stock soln every 6-12 months) directly to
the culture media (~1.8%) for 5 minutes at RT.  Followed by centrifugation
for 5 min cells are resuspended in excess volume 1 x PBS and stored at 4
C.  An increase in fluorescence can often be observed as more GFP can fold
at 4 C O/N.  These cells can then be stained with DAPI for cell-cycle or
with Ab for a second label.

Happy GFPing,

Mike Moser                                            Tel: 206-543-6585
UW Department of Pathology                            FAX: 206-543-3967
Box 357705                                       moser at u.washington.edu
Seattle, WA  98195                 http://weber.u.washington.edu/~moser

On 7 Nov 1997 Jacqueline_Glynn at cc.chiron.com wrote:

> I was wondereing whether anyone knows a protocol for fixing EGFP-expressing
> cells for flow cytometry? Will the standard 1% paraformaldehyde fixation work?
> 
> Jackie Glynn
> Jacqueline_Glynn at cc.chiron.com
> 
> 




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