The only stimulus we have gotten to work for IL-10 in fresh T cells is PMA/iono at 6 hours. We have had good results with the 9D7 clone, preferably conjugated to PE or APC. The frequency of IL-10 pos CD4 T cells is typically <<1%. Calman > ----- Original Message ----- > From: Joost Schuitemaker > To: Cytometry Mailing List > Sent: Thursday, May 10, 2001 3:29 AM > Subject: Intracellular staining of IL10 > > > > Dear friends, > > For a few years now intracellular staining of cytokines is performed > almost routinely in some labs (including ours), but there are still some > cytokines that give difficulties. For instance TGF's, IL18 and IL10. And > especially this last one is more and more investigated by many people. > Detection (intracellularly) of IL10 production by monocytes after > stimulation with LPS or CD40L is no problem. But what about lymphocytes? > We would like to look at this cytokine in HUMAN T cells. And when > possible in combination with other cytokines. Has anyone ideas concerning > timepoint, kind of stimulus and antibody to use? Litt. is not conclusive > and detection in ELISA is much earlier than seeing 0.0X % IL10 positive > cells (with the same stimulation) in intracellular staining. Can someone > help us? > > Kind regards, > > Joost Schuitemaker > > ______________________________________________________________ > > J.H.N. Schuitemaker > Senior Research Technician > Dept. Cell Biology & Histology > Cellular Immunology Group > AMC, University of Amsterdam > Meibergdreef 9, L3-351 PObox 22700 > 1105 AZ Amsterdam 1100 DE Amsterdam > The Netherlands > > Interested? VISIT: > (intranet) http://m3.amc.uva.nl/celbiologie > (internet) http://www.amc.uva.nl/ and > (internet) http://home.wanadoo.nl/flowcytometry/ > > Telephone: +31 (0)20 5664960 > Fax: +31 (0)20 6974156 > > E-mail (privé): J.H.N.Schuitemaker@consunet.nl > > > > >
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