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>>
