From: medbury@renal.wsahs.nsw.gov.au
Date: Fri May 29 1998 - 21:39:33 EST
Hi flow-ers I am starting to dabble with intracellular cytokine detection by flow cyto in mouse lymphocytes. To start with I just want to look at CD3 and IL-2 to get the method working. I am following the Pharmingen methods. I am just going to stimulate the cells with a mitogen first does anyone know the best conditions?? I tried PMA 10ng/ml, ionomycin 1uM, 2-ME 100uM and monensin 1um for 16hrs ie overnight, but I couldnt detect IL-2, even when I gated on the CD3 population. thanks for any replies most appreciated Heather Heather Medbury PhD Department of Surgery Westmead Hospital Westmead, NSW, 2145 Australia ph (612) 9845 7680 fx (612) 9893 7440
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