FLICA 660 Poly Caspase Assay Kit
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.
FLICA 660 Poly Caspase Assay Kit
Caspases play important roles in apoptosis and inflammation. ICT’s FLICA assay kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells. The FLICA 660 Poly Caspase probe allows researchers to assess caspase activation.
The FLICA reagent 660-VAD-FMK enters each cell and irreversibly binds to activated caspases. Because the 660-VAD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound 660-VAD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining far red fluorescent signal is a direct measure of the active caspase enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by fluorescence microscopy or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 16 hours using the fixative. Unfixed samples may also be analyzed with Hoechst stain to detect changes in nuclear morphology.
660-VAD-FMK
Poly caspases
660 nm / 690 nm
Flow cytometry, Fluorescence microscope
Cell culture
2-8°C
Domestic: Overnight Delivery; International: Priority Shipping
United States
- 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