Dear flowers, I'd like to detect apoptosis of murine dendritic cells and T-cells using anexinV / PI staining (using a BenderMedsystem kit). Cells are co-cultured for 24h. For cell discrimination, the surface should be stained with CD11c or CD8/CD4 APC (for detection on standard two-laser Calibur). Anybody did this before? Should I stain first with the APC-labelled antibodies? Or first Annexin? And wash thereafter? Or both together? And PI before analysis? Any suggestions? Hints? Thanks
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