From: Simon Monard (smonard@trudeauinstitute.org)
Date: Thu Dec 12 2002 - 19:48:36 EST
Hey Jose We did a similar thing a few years back. We would do our surface staining, incubate with BD FACS lysing solution for 10 or 15 minutes wash a couple of times then stain with the Ki67 antibody. It worked OK. The intensity of the Ki67 could be improved a bit by adding a bit of detergent like NP 40, I think it was 0.05% at the same time as the Ki67 antibody. Playing about with the detergent type/concentration would probably help. You will lose most surface staiing if you use ethanol plus you will lose a lot of cells so probably distorting your results. Surface staining then fixing with 0.5% formaldehyde (decent EM grade) for 5-15 minutes before using ethanol might work, its always a compromise between retaining surface staining and doing a good job of permiabilizing. Good luck Simon Monard Trudeau Institute New York >>> "Jose Benito" <jbenito1@hotmail.com> 12/12/02 18:48 PM >>>
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