I also have used the same clone of IFN-g, as well as the Pharmingen recomended clone for IL-4, IL-6, IL-8, MCP-1, GM-CSF and MIP-1a.......IL-4 and IFN-g require PMA (50 ng/ml) and calcium ionophore (1 ug/ml) and monensin for 24 hours. IL-6, IL-8, MCP-1 and MIP-1a require LPS 10 ng/ml and monensin 24 hours , GM-CSF requires I-2 and IL-4 for 6 days, then ionomycin and PMA and monensin for 24 hours. Specifics are in a paper published by Pharmingen last October, although I adjusted the GM-CSF activation from what was in that paper. Patricia L. Echeagaray <$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$><$> Manager, Flow Cytometry Services Southern Research Institute 431 Aviation Way Frederick, Maryland 21701 Voice: 301-694-3232 FAX: 301-694-7223 E-mail: echeagaray@sri.org > -----Original Message----- > From: Gillis_Otten@cc.chiron.com [SMTP:Gillis_Otten@cc.chiron.com] > Sent: Wednesday, March 29, 2000 3:50 PM > To: Cytometry Mailing List > Subject: Re: intracellular cytokine staining in non-human primates > > > For TNFalpha we use MAb11 and for IFNgamma we use 4S.B3 to stain > rhesus. (These also work for baboon). Both reagents come from > Pharmingen. We have used FITC and APC conjugates for TNFalpha with > very similar results. We have used only the PE-conjugated 4S.B3 for > IFNgamma. We have not tried either IL2 or IL4 staining. > > Gib Otten > Chiron Corp > Emeryville, CA > > > ______________________________ Reply Separator > _________________________________ > Subject: intracellular cytokine staining in non-human primates > Author: Laura Vitale <lvitale@injersey.infi.net> at SMTP > Date: 3/28/00 11:26 AM > > > Hi! > Can anyone give me information related to intracellular cytokine > detection by flow cytometry in rhesus/macaque T cells? I'm specifically > looking for information regarding human antibodies that cross-react > (IL-2, TNF-a, IL-4 and g-IFN) and possible protocols for staining. > > Thanks, > Laura Vitale > Medarex, Inc > Annandale, NJ > 908-713-6000 x338
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