Hi flowers, I am doing some T helper differentiation experiments and am just wondering about some specifics that people are doing. If people magnetically select CD4+ T cells and then only culture these cells, for like 5 day later staining, do people restain for CD4 and then look at e.g IL17, IFN-Y production? Or do you only stain for these cytokines>?? And just choose everything that falls in the lymphocyte gate. What numbers of t cells are seeded to get good %'s when running flow. Example seeded 100000 cells / well. 200uL volume, is this usually enough? Because I am having trouble getting many events after all the staining, intracellular staining, etc etc. Any other ideas/useful heads upss would be greatly appreciated. Thanks --------------------------------------------------- Suat Dervish PhD Student The Sutton Arthritis Research Laboratories Royal North Shore Hospital NSW, Australia Phone:9926 6251 Email: sdervish@med.usyd.edu.au ---------------------------------------------------Received on Fri Jun 27 15:58:00 2008
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