FAM FLICA Caspase 1
97 kit98 kit

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 or tissue samples. Analyze the green fluorescent signal using fluorescence microscopy, a fluorescent plate reader, or by flow cytometry.

Catalog # Size Quantity Price
97 25 tests $205.00
98 100 tests $575.00
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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 fluorescent plate reader, fluorescence microscopy, or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 24 hours using the fixative. Unfixed samples may also be analyzed with propidium iodide or Hoechst stain to detect necrosis or changes in nuclear morphology respectively.

1. Prepare samples and controls
2. Dilute 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. Wash and spin cells three times.
8. If desired, label with additional stains, such as Hoechst, propidium iodide, 7-AAD, or an antibody.
9. If desired, fix or embed 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 frozen tissues or 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, tissue
• Storage: 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping

Kit 97: 25 tests
• FLICA Caspase-1 Reagent (FAM-YVAD-FMK), 1 vial
• 10X Apoptosis Wash Buffer, 15 mL
• Fixative 6 mL
• Hoechst 33342 Stain, 1 mL
• Propidium Iodide Stain, 1 mL

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