Hi David, Considering a good number of markers will get up- or downregulated on stimulation, it would be useful to let us know just what the purpose of the stimulation is in what seems to be a straightforward phenotyping experiment. Otherwise, you can use any number of lectins or antiCD3/CD28 beads in whole blood, but please beware of cells clustering together after stimulation. Why do you think you'd lose rare populations in Ficoll? Perhaps an erythro lysis step would be a simple alternative but again, everything hinges on what you're trying to accomplish here. Guy Guy Hermans, PhD Ablynx N.V. Technologiepark 4 9052 Zwijnaarde Belgium Phone : +32 9 261 06 23 Fax : +32 9 261 06 27 _______________________________________________ -----Original Message----- From: Bruno, David (NIH/NIDDK) [mailto:brunod@niddk.nih.gov] Sent: Friday, December 03, 2004 8:51 PM To: cyto-inbox Subject: PBMC Stimulation Dear Flowers, I am looking for the cleanest way to stimulate PBMC. I would like to avoid ficoll because I do not want loose rare cell populations. I would also like to avoid anything that would prevent phenotyping with CD3, 4, 8, 45RA, 62L etc... Thanks, David A. Bruno, MD Research Fellow National Institutes of Health NIDDK/ Transplantation Branch CRC 5-5832 301-451-3347 ----------------------------------------------------------------------- THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE. If the reader of this E-mail message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately at ablynx@ablynx.com. Thank you for your co-operation. -----------------------------------------------------------------------Received on Tue Dec 7 14:18:01 2004
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