From: Guy Hermans (Guy.Hermans@ablynx.com)
Date: Wed Dec 17 2003 - 03:38:02 EST
The Miltenyi system crosslinks anti-cytokine capture antibody to an anti-CD45 mAb. I didn't try, but I suppose you buy an ELISA "matched pair" anti-cytokine antibodies, the detection mAb being biotinylated, and crosslink the capture clone to anti-CD45 or a monoclonal more specific for your cell. Pierce sells kit for that kind of thing. I guess you'd need an FPLC or HPLC system to formally check for efficient crosslinking, or purify the crosslinked fraction away from uncrosslinked anti-surface antigen monoclonal (competitive binder!). Reactivity of the new conjugate towards your antigens after coupling needs to be checked in FACS and ELISA, respectively, but that's not too much work either. Regards, 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: Joanne Lannigan [mailto:joannelannigan@virginia.edu] Sent: Tuesday, December 16, 2003 4:57 PM To: cyto-inbox Subject: Sorting live cytokine producing cells I need to be able to sort live IL17 producing T cells. Does anyone have any ideas on how to achieve this? I thought of the cytokine capture assay but I could not find anything for IL17. Any suggestions? Thanks- Joanne Lannigan, MS Director, Flow Cytometry Core Facility University of Virginia Jordan Hall, Room 7067 P.O. Box 800734 Charlottesville, VA 22908-0734 Office: 434-924-0274 Lab: 434-243-2695 Fax: 434-982-1071 email: joannelannigan@virginia.edu ----------------------------------------------------------------------- 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. -----------------------------------------------------------------------
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