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FAM FLICA® Caspase-1 (WEHD) Assay Kit

$211.00$592.00

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

Catalog # Product Size Quantity Price
9161 25 Tests $211.00
9162 100 Tests $592.00

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

ICT’s FAM-FLICA Caspase-1 (WEHD) kit (#9162) was used to detect pyroptosis in THP-1 monocytes. Cells were labeled with FAM-WEHD-FMK, treated with a negative control (DMSO) or LPS (1 µg/mL, 3 hours) followed by Nigericin (20 µM, 30 minutes) to induce pyroptosis. Following treatment, cells were washed and resuspended in PBS + 0.5% BSA. Cells were then stained with Hoechst 33342, fixed, and viewed using a Logos iRiS Digital Cell Imaging System equipped with a EGFP (Ex 470/30, Em 530/50) and a DAPI (Ex 375/28, Em 460/50) LED filter cubes at 20X. Increased levels of green FAM-FLICA staining (leftmost images) were seen in the LPS + Nigericin-treated cells (upper row of images) compared to DMSO mock-treated cells (lower row of images). Hoechst 33342 staining (middle images) and overlays of both the EGFP and DAPI channels (rightmost images) are also shown.

WEHD Data Tab Image

• Reagent name: FAM-WEHD-FMK
• Target: Caspase-1
• Excitation/Emission: 492 nm / 520 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 9161: 25 Tests
• FLICA Caspase-1 Reagent (FAM-WEHD-FMK), 1 vial, #6708
• 10X Apoptosis Wash Buffer, 15 mL, #635
• Fixative, 6 mL, #636
• Propidium Iodide, 1 mL, #638
• Hoechst 33342, 1 mL, #639
• Kit Manual

Kit 9162: 100 Tests
• FLICA Caspase-1 Reagent (FAM-WEHD-FMK), 4 vials, #6708
• 10X Apoptosis Wash Buffer, 60 mL, #634
• Fixative, 6 mL, #636
• Propidium Iodide, 1 mL, #638
• Hoechst 33342, 1 mL, #639
• Kit Manual