JC-1 Fluorescent Mitochondrial Membrane Depolarization Assay

1. Prepare samples and controls., 2. Dilute Apoptosis Wash Buffer 1:10 with diH20., 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. For example, to stain at 1:30, add 10 µL to 290 µL of cultured cells. To stain at 1:60, add 5 µL to 295 µL of cultured cells., 6. Incubate approximately 1 hour., 7. Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells., 8. If desired, label with additional stains, such as Hoechst, 7-AAD, or an antibody., 9. If desired, fix or embed cells., 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. SR-FLICA excites at 550-580 nm and emits at 590-600 nm.