Annexin V-FITC Apoptosis Assay Kit

1. Prepare samples and controls, 2. Dilute 10X Apoptosis Wash Buffer 1:10 with diH2O., 3. Reconstitute FLICA with 50 µL DMSO., 4. Dilute FLICA 1:5 by adding 200 µL PBS., 5. Add diluted FLICA to each sample at 1:30-1:60 (e.g. spike at 1:30 by adding 10 µL to 290 µL cultured cells)., 6. Incubate approximately 1 hour., 7. Wash and spin cells three times., 8. If desired, label with additional stains, such as Hoechst, DAPI, or an antibody., 9. If desired, fix cells., 10. Analyze with a fluorescence microscope or flow cytometer. FLICA 660 excites at 660 nm and emits at 680-690 nm.