MitoPT JC-1 Assay Kit (400 tests)

1. Prepare samples and controls. 2. Dilute 10X 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 (e.g. add 10 μL to 290 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, Propidium Iodide, 7-AAD, or an antibody. 9. If desired, fix cells. 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM FLICA excites at 492 nm and emits at 520 nm. If working with adherent cells, please see the manual for additional protocols.