Detect active caspase enzymes with the FLICA 660 Poly Caspase Assay Kit. This in vitro assay employs the fluorescent inhibitor probe 660-VAD-FMK to label active caspase enzymes in living cells. Analyze the fluorescent signal using fluorescence microscopy 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 1:30-1:60 (e.g. spike at 1:30 by adding 10 µL to 290 µL cultured cells).
- Incubate approximately 1 hour.
- Wash and spin cells three times.
- If desired, label with additional stains, such as Hoechst, DAPI, or an antibody.
- If desired, fix cells.
- Analyze with a fluorescence microscope or flow cytometer. FLICA 660 excites at 660 nm and emits at 680-690 nm.
Elsherbini A, Kirov AS, Dinkins MB, Wang G, Qin H, Zhu Z, Tripathi P, Crivelli SM, Bieberich E. Association of Aβ with ceramide-enriched astrosomes mediates Aβ neurotoxicity. Acta Neuropathol Commun. 2020 Apr 28;8(1):60. doi: 10.1186/s40478-020-00931-8. Full Text