FAM-FLICA® Caspase-1 Assay Kit

$205.00$575.00

Detect caspase-1 activity with the FAM FLICA Caspase-1 Assay Kit. This in vitro assay employs the fluorescent inhibitor probe FAM-YVAD-FMK to label active caspase-1 enzyme in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.

Catalog # Size Quantity Price
97 25 tests $205.00
98 100 tests $575.00
Categories: ,

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-1 probe allows researchers to assess caspase-1 activation.

The FLICA reagent FAM-YVAD-FMK enters each cell and irreversibly binds to activated caspase-1. Because the FAM-YVAD-FMK FLICA reagent becomes covalently coupled to the active enzyme, it is retained within the cell, while any unbound FAM-YVAD-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-1 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.

1. Prepare samples and controls
2. Dilute 10X Apoptosis Wash Buffer 1:10 with diH20.
3. Reconstitute FLICA with 50 μL DMSO.
4. Dilute FLICA 1:5 by adding 200 μL PBS.
5. Add diluted FLICA to each sample at 1:30 (e.g., add 10 μL to 290 μL of cultured cells).
6. Incubate approximately 1 hour.
7. Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells.
8. If desired, label with additional stains, such as Hoechst, Propidium Iodide, 7-AAD, or an antibody.
9. If desired, fix cells.
10. 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.

Suspension cells were treated with an apoptosis-inducing agent or DMSO, a negative control, for 4 hours, washed twice, then incubated with ICT’s green caspase-1 inhibitor probe, FAM-YVAD-FMK, for 1 hour and examined under a fluorescence microscope. The top images reveal several experimental cells, all of which fluoresce green, therefore they have active caspase-1. The lower brightfield image at right reveals many control cells in the field of view, however the corresponding fluorescence image is dark (lower left); none of these cells have active caspase-1 (Dr. Brian W. Lee, ICT).caspase1data_1

• Reagent name: FAM-YVAD-FMK
• Target: Caspase-1
• Excitation/Emission: 488 nm / 530 nm
• Method of Analysis: Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
• Types of Samples: Cell culture
• Storage: 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping

Kit 97: 25 tests
• FLICA Caspase-1 Reagent (FAM-YVAD-FMK), 1 vial, #655
• 10X Apoptosis Wash Buffer, 15 mL, #635
• Fixative, 6 mL, #636
• Propidium Iodide, 1 mL, #638
• Hoechst 33342, 1 mL, #639
• Kit Manual

Kit 98: 100 tests
• FLICA Caspase-1 Reagent (FAM-YVAD-FMK), 4 vials, #655
• 10X Apoptosis Wash Buffer, 60 mL, #634
• Fixative, 6 mL, #636
• Propidium Iodide, 1 mL, #638
• Hoechst 33342, 1 mL, #639
• Kit Manual