--- On Tue, 5 Jan 1999 13:41:37 PST frances gibson <fgibson@sghms.ac.uk> wrote: > >Thinking about positive controls for surface Fas and Fas-L expression >by flow. >Am I right in thinking that the line Hut78 constitutively expresses >Fas-L and Jurkats express Fas. >Also what will upregulate Fas-L on PBMCs ? PHA, PMA and/or ionomycin ? >Are these reagents acting on different cell types ie. T cells versus >monocytes ? Will the same reagents upregulate Fas expression ? > Yes, Jurkat express constitutively high levels of Fas. Anumber of stimuli have been reported to upregulate FasL including superAg, anti CD3, PMA+iono, CD4 crosslinking, con A, IL-2. Fas expression on PBMC can be upregulated by CD4 xlink, TNF, IFN. You will need to use an inhibitor to prevent FasL from being shed off the membrane. I'm not familliar with HUT78, we have used FasL transfectants as positive controls. You can find more specific info in these references: J Immunol 158: 2456-2463, 1997 Blood, 84: 2622-2631, 1994 Cell Immunol 179: 153-164, 1997 EMBO J 14: 1129-1135, 1995 J EXp Med 181: 71-77, 1995 J Immunol 154: 3806-3813, 1995 J Infect Dis 178: 1030-1039, 1998 Tom -------------------------------------------------------- Thomas W. Mc Closkey, Ph. D. Director, Flow Cytometry North Shore University Hospital Biomedical Research Center 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office]; 516-562-1135/4641 [lab] fax: 516-562-2866 1/6/99 5:49:28 PM E-mail: thomasm@nshs.edu --------------------------------------------------------
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