Date: Wed Jun 07 2006 - 11:19:00 EDT
Hi Jose, I fail to see what keeps you from using a negative selection CD8 kit or multi-sort kit on the PBMC first and run an IFNg secretion assay positive sort later. Both should be doable both before or after T-cell stimulation, but since I don't know if CD8 gets downregulated after stimulation you would do well to [1] check CD8 expression before and after stimulation of your sample prior to any sorting experiments or [2] simply stimulate with a CD8- population of APC's or antiCD3/CD28 as the case may be after CD8+ T-cell isolation. best of luck, Guy Next generation therapeutic <http://www.ablynx.com/> antibodies Guy Hermans, PhD Senior Scientist Ablynx NV Technologiepark 4 B-9052 Zwijnaarde Belgium guy.hermans@ablynx.com tel: fax: mobile: +32 (0)9 261 06 57 +32 (0)9 261 06 27 +32 (0)486 788 551 <https://www.plaxo.com/add_me?u=30065269879&v0=994426&k0=2009290972> Add me to your address book... <http://www.plaxo.com/signature> Want a signature like this? -----Original Message----- From: Jose Benito [mailto:jbenito1@hotmail.com] Sent: Thursday, May 18, 2006 12:53 PM To: cyto-inbox Subject: separation of IFN-producing viable cells Dear flowers, we are interested in the sorting of Ag-specific IFN-producing CD8+ cells. We are considering the Miltentyi IFN-gamma secretion and purification assay. However, if I understand correctly, with this assay you can only separate every PBMC able to produce IFN, and can not discriminate between CD8+ and other cells (for example NK cells). In previous experiments we have found that in some samples a large proportion od CD8- cells (most likely NK cells) produce IFN in response to Ag. So how can we sort only CD8+IFN+ cells using the Miltenyi assay? Best regards Jose M Benito Madrid ----------------------------------------------------------------------- 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. -----------------------------------------------------------------------
