This in vitro assay employs the far-red fluorescent inhibitor probe 660-VAD-FMK to label active caspase enzymes in living cells. Analyze the fluorescent signal using fluorescence microscopy or flow cytometry.
Proben:
Cell culture
Anwendungsbeschreibung:
Application Notes: 1. Prepare samples and controls., 2. Dilute 10X Cellular 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 (e.g. spike at 1:30 by adding 10 µL to 290 µL cultured cells)., 6. Incubate approximately 1 hour., 7. Remove media and wash cells 3 times: add 1X Cellular Wash Buffer and spin cells., 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
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