Hi Jose, We were having the same problem when we started out on our quest to incorporate CD107 in our assays. I have the following suggestions. 1) If you are using thawed PBMCs , try to rest them for a day before stimulating them with the peptide. 2) Did you get your compensations right. what fluorochrome are you using your CD107 with? what are the other colors in the assay. 3) try using a viability marker and exclude the dead cells as they bind CD107 non specifically. 4) are you using this as part of a cytokine assay (looking for IFN-g etc?) Thanks Vijay Nandakumar, Cellular and Molecular Immunology Laboratory Mayo Clinic 200, 1st street SW, Rochester, MN, 55905 ________________________________ From: owner-cyto-sendout@flowcyt.cyto.purdue.edu [mailto:owner-cyto-sendout@flowcyt.cyto.purdue.edu] On Behalf Of Jose Benito Sent: Tuesday, December 12, 2006 3:06 AM To: cyto-inbox Subject: CD107 Hi eveyone, I am working with CD107 antibody as a marker of CTL degranulation, in combination with several other markers. I am having problems with unstimulated controls, usually finding high values of CD8 cells positive for CD107. Is there any way to solve this problem? Even if I simply substract the unstimulated control from the stimulated one, I find unusually high values for CD8+CD107+ peptide-specific cells. So I suspect this is an artifact rather than real values. Many thanks José M Benito Infectious Diseases Hospital Carlos III Madrid Spain ________________________________ FREE pop-up blocking with the new MSN Toolbar MSN Toolbar <http://g.msn.com/8HMAEN/2752??PS=47575> Get it now!Received on Wed Dec 13 13:38:01 2006
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