FAM-FLICA® Caspase-3/7 Assay Kit
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.
FAM-FLICA® Caspase-3/7 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 and tissues. The FAM FLICA Caspase-3/7 assay probe allows researchers to assess caspase 3 and 7 activation.
The FLICA reagent FAM-DEVD-FMK enters each cell and irreversibly binds to active caspases 3 and 7. Because the FAM-DEVD-FMK FLICA reagent becomes covalently coupled to the active enzymes, it is retained within the cell, while any unbound FAM-DEVD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining green fluorescent signal is a direct measure of the active caspase 3 and 7 enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by a fluorescence plate reader, fluorescence microscopy, or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 16 hours using the fixative included in the kit. Unfixed samples may also be analyzed with Propidium Iodide or Hoechst 33342 to detect necrosis or changes in nuclear morphology, respectively.
FAM-DEVD-FMK
Caspase-3/7
488 nm / 530 nm
Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
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 (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.
Kit 93: 25 Tests
FLICA Caspase-3/7 Reagent (FAM-DEVD-FMK), 1 vial, #653
10X Apoptosis Wash Buffer, 15 mL, #635
Fixative, 6 mL, #636
Propidium Iodide, 1 mL, #638
Hoechst 33342, 1 mL, #639
Kit Manual
Kit 94: 100 Tests
FLICA Caspase-3/7 Reagent (FAM-DEVD-FMK), 4 vials, #653
10X Apoptosis Wash Buffer, 60 mL, #634
Fixative, 6 mL, #636
Propidium Iodide, 1 mL, #638
Hoechst 33342, 1 mL, #639
Kit Manual
Product Specific References
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