Detect caspase-3/7 activity with the FLICA Caspase-3/7 Assay Kit. This in vitro assay employs the fluorescent inhibitor probe FAM-DEVD-FMK to label active caspase 3 and 7 enzymes in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.
- Prepare samples and controls
- Dilute 10X Apoptosis Wash Buffer 1:10 with diH20.
- Reconstitute FLICA with 50 µL DMSO.
- Dilute FLICA 1:5 by adding 200 µL PBS.
- Add diluted FLICA to each sample at 1:30 (e.g., add 10 µL to 290 µL of cultured cells).
- Incubate approximately 1 hour.
- Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells.
- If desired, label with additional stains, such as Hoechst, Propidium Iodide, 7-AAD, or an antibody.
- If desired, fix cells.
- 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.
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