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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