Hello Christiane, We got a nice protocol from Kris Thielemans' lab in Brussels where the PBMC's are spun down at relatively low g. PBMC are resuspended in about 35 ml of PBS/2mM EDTA (R.Tē) and washed 3 times 15 min/800rpm/20ēC. The first 1 or 2 times the supernatant will appear rich in cells: don't be afraid to discard it! (check under a microscope, it's all PLTs). After the 3rd wash, the supernatant should look crystal-clear, otherwise continue washing the same way. Good luck. Karim "Dr. Christiane Hollmann" wrote: > 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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