SR-FLICA® Caspase-9 Assay Kit
Detect caspase-9 activity with the SR-FLICA Caspase-9 Assay Kit. This in vitro assay employs the fluorescent inhibitor probe SR-LEHD-FMK to label active caspase-9 enzyme in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.
SR-FLICA® Caspase-9 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 FLICA reagent SR-LEHD-FMK enters each cell and irreversibly binds to activated caspase-9. Because the SR-LEHD-FMK FLICA reagent becomes covalently coupled to the active enzyme, it is retained within the cell, while any unbound SR-LEHD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining red fluorescent signal is a direct measure of the active caspase-9 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 Hoechst 33342 to detect changes in nuclear morphology.
SR-LEHD-FMK
Caspase-9
550-580 nm / 590-600 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 – 1:60. For example, to stain at 1:30, add 10 μL to 290 μL of cultured cells. To stain at 1:60, add 5 μL to 295 μL of cultured cells.
- Incubate approximately 1 hour.
- Remove media and wash cells 3 times: add 1X Apoptosis Wash Buff er and spin cells.
- If desired, label with additional stains, such as Hoechst, 7-AAD, or an antibody.
- If desired, fix or embed cells.
- Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. SR-FLICA excites at 550-580 nm and emits at 590-600 nm.
Kit 960: 25 Tests
FLICA Caspase-9 Reagent (SR-LEHD-FMK), 1 vial, #6145
10X Apoptosis Wash Buffer, 15 mL, #635
Fixative, 6 mL, #636
Hoechst 33342, 1 mL, #639
Kit Manual
Kit 961: 100 Tests
FLICA Caspase-9 Reagent (SR-LEHD-FMK), 4 vials, #6145
10X Apoptosis Wash Buffer, 60 mL, #634
Fixative, 6 mL, #636
Hoechst 33342, 1 mL, #639
Kit Manual