Hello! I'm interested in surface staining platelets. I have a protocol (from Current Protocols in Cytometry) for staining whole blood, which uses thiazole orange coupled with a platelet-specific monoclonal antibody. I'd rather isolate the platelets from whole blood first, then stain them (so that I can avoid having to do multicolor analysis). It seems to be relatively simple to isolate platelets from plasma, but I've had trouble with them forming clumps. I used a wash buffer of PBS, 1% FBS, and 15mM EDTA. Possibly my concentration of EDTA was too low? Has anyone worked with platelets who can offer a few hints?? Thanks! Jeannine Navratil University of Pittsburgh Arthritis Institute
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