Hi Lisa, Assuming viability was as good as the purity, you are doing to right thing. Most likely, the source of cells was contaminated with large numbers of activated effector cells, which have the same phenotype as "suppressors". There are a few putative markers for suppressor cells out there, some of which require permeabilizing your cells (hence impossible to validate on suppression behaviour afterwards ;-) and the true value of virtually all these markers is as debatable as that of CD25 itself. Therefore, your best solution for now would be to get another type of input sample from the customer. Guy ------------------------------------ Ablynx NV Guy Hermans, PhD Senior Scientist guy.hermans@ablynx.com Technologiepark 4 B-9052 Zwijnaarde Belgium tel: +32 (0)9 261 06 57 fax: +32 (0)9 261 06 27 mobile: +32 (0)486 788 551 ------------------------------------ -----Original Message----- From: lixin86 [mailto:lixin86@interchange.ubc.ca] Sent: Thursday, July 20, 2006 9:31 PM To: cyto-inbox Subject: Mouse CD4+CD25+ Regulatory T cells sorting? Hi, I Sorted Mouse CD4+CD25+ Regulatory T cells from total spleenocytes on BD FACSAria. My customer asked me to sort around 5% top CD25+ of total CD4+ T cells. After sorting by 70 nozzle on either 70 psi or 35 psi, the purity is around 98-99%. However, the sorted population had no suppression function. Can anyone tell me your kind advice? Lisa -- ******************************** BC Research Institute, UBC Room 212, 950 W 28th Ave. Vancouver, BC V5Z 4H4 Canada Tel: 604-875-2000 ext 5987 Fax: 604-875 3597 ----------------------------------------------------------------------- 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. "NANOBODY" and "NANOCLONE" are registered trademarks of Ablynx N.V. -----------------------------------------------------------------------Received on Mon Jul 24 13:38:00 2006
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