Christiane, I have a couple of suggestions: First, we typically wash our PBMCs repeatedly with HBSS+1%BSA, spinning at around 220 xg. We repeat these washes until the supernatant is clear. Time consuming, but effective. A new technique you might try utilizes products from StemCell Technolgies in Vancouver. It's a product line called RosetteSep. They use antibody dimers, where the tail of the dimer is an anti-glycophorin antibody, to cause unwanted cells to aggregate and pass through the Ficoll gradient. I think they include an anti-platelet antibody in their product list, but if not, they can make you a custom cocktail for a reasonable price. If you want quick results and cost is not an object, give it a shot. http://www.stemcell.com/ Best of luck. kb p.s. Of course, I would NEVER suggest you use anything for cell separation besides a MoFlo......... :-) Happy holidays. ------ Keith Bahjat, Ph.D. Applications Engineering Manager Cytomation, Inc Fort Collins, Colorado Phone: (970) 226-2200 x223 Fax: (970) 226-0107 keithb@cytomation.com -----Original Message----- From: Dr. Christiane Hollmann [mailto:ch@adnagen.com] Sent: Wednesday, November 21, 2001 1:51 AM To: cyto-inbox Subject: depletion of platelets Dear Flowers, I have a problem to get rid of remaining platelets after MNC preparation via Ficoll gradient. Up to now we centrifuge the MNC through FCS or BSA. Are their any protocols to increase the efficiency of platelet depletion? Please let me know! Regards Christiane
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