Re: [CD8 after EtOH fixation]

From: Simon Monard (smonard@trudeauinstitute.org)
Date: Wed Jan 23 2002 - 11:50:30 EST


You can try fixing with 1% or 0.5% formaldehyde for 10 or15 mins then washing and then
doing your ethanol fixation. You may have to play about with duration of formaldehyde
fixation. Its always a bit of a compromize between good surface staining and a decent
DNA profile.


Simon Monard
FACS Lab Manager
Trudeau Institute
Saranac Lake
NY12983

Ph 518 891 3080 X352


>>> Philippe Pognonec <Philippe.Pognonec@unice.fr> - 1/22/02 10:01 AM >>>

Hi,
We want to label CD8alpha expressing cells with an FITC conjugated Ab. This Ab works
great on T cells (Pharmingen, clone
53-6.7). But we need to have our cells EtOH fixed for further DNA content analysis
with PI, instead of the usual formaldehyde
procedure, which is incompatible with PI DNA content analysis. Up to now, our first
tests are not very promising. Does anybody
out there have some experience on that point, and good advice?
Thank you in advance!
Philippe
PS: by the way, if someone knows where we can get the 53-6.7 clone, I am sure our lab
budget would appreciate the tip...

____________________________________________

Philippe Pognonec, Ph.D.
Transcriptional Regulation and Differentiation
Centre de Biochimie
Parc Valrose
Universite de Nice
06108 Nice cedex 2
France
Tel/fax: (33) 492 07 64 13
____________________________________________



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