If, after fixation in formaldehyde, you suspend the cells in PBS and heat at 75 degrees Centigrate for 1 hr , before reacting with antibody then with PI, the staining properties of the cells with PI are largely restored. Not as good as EtOH, but might be worth a try. ----any port in a storm. -----Original Message----- From: Philippe Pognonec [mailto:Philippe.Pognonec@unice.fr] Sent: Tuesday, January 22, 2002 10:01 AM To: cyto-inbox Subject: [CD8 after EtOH fixation] 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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