Pyroptosis FAM Caspase-1
|Pyroptosis FAM Caspase-1 Kit utilizes the popular FLICA technology to detect caspase-1 activation. FLICA probes are cell permeant non-cytotoxic. This kits contain the caspase-1 inhibitor reagent YVAD-FMK, which has the preferred binding sequence for caspase-1, Tyr-Val-Ala-Asp (YVAD) (Chapman, 1992). This preferred caspase-1 binding sequence is labeled with either FAM a green fluorescent dye and linked to a fluoromethyl ketone (FMK) reactive entity. Caspase-1 will not cleave the FLICA inhibitor probe; instead, it forms an irreversible covalent bond with the FMK group on the reagent and becomes inhibited from further enzymatic activity.|
- Reagents in the Kit
- 4 vials of FAM-YVAD-FMK Reagent - lyophilized
1 vial Nigericin - lyophilized
10X Wash Buffer, 60 ml
Fixative, 6 mL
Hoechst Stain, 1 ml.
- Test Principle
- To use FLICA, add directly to the cell culture medium, incubate, and wash. FLICA is cell-permeant and will efficiently diffuse in and out of all cells. If there is an active caspase-1 enzyme inside the cell, it will covalently bind with YVAD-FMK and retain the fluorescent signal within the cell. Unbound FLICA will diffuse out of the cell during the subsequent wash steps. Therefore, positive cells will retain a higher concentration of FLICA and fluoresce brighter than negative cells. There is no interference from pro-caspases or inactive forms of the enzymes. After labeling with FLICA, cells can be counter-stained with other reagents and fixed or frozen. Cells labeled with YVAD-FMK can be counter-stained with reagents such as the red live/dead stains Propidium Iodide and 7-AAD. Nuclear morphology may be concurrently observed using Hoechst 33342 (included in the kit), a blue DNA-binding dye. Cells can be viewed through a fluorescence microscope or flow cytometer.
- Max Ex/Em
Fluorophore Excitation Max (nm) Emission Max (nm) FAM 494 520
- For research purposes only.
- Guaranteed until date of expiry. Please see product label.
- FLICA is a trademark of Immunochemistry Technologies, LLC.
|Application Name||Verified||Min Dilution||Max Dilution|
|Flow Cytometry||Refer to Instructions for Use|
- Instructions For Use
- Instructions for use can be found at https://www.bio-rad-antibodies.com/static/uploads/ifu/ict9145-6.pdf
Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"
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