FAM FLICA® Caspase 1 Assay Kit
Detect caspase-1 activity with the FLICA® Caspase 1 Assay Kit. This in vitro assay employs the green fluorescent inhibitor probe FAM-YVAD-FMK to label active caspase 1 enzyme in living cells or tissue samples. Analyze the fluorescent signal using fluorescence microscopy, a fluorescent plate reader, or by flow cytometry. Caspase 1 (ICE):Members of the mammalian caspase family of cysteinyl aspartate-specific proteases play disti…
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Detect caspase-1 activity with the FLICA® Caspase 1 Assay Kit. This in vitro assay employs the green fluorescent inhibitor probe FAM-YVAD-FMK to label active caspase 1 enzyme in living cells or tissue samples. Analyze the fluorescent signal using fluorescence microscopy, a fluorescent plate reader, or by flow cytometry.
Caspase 1 (ICE):
Members of the mammalian caspase family of cysteinyl aspartate-specific proteases play distinct roles in apoptosis and inflammation. Originally identified as Interleukin -1β Converting Enzyme or ICE, caspase 1 along with caspases 4, 5, and 12 comprise the inflammatory subfamily of caspase enzymes. In rodents, caspase 11 is also an inflammatory caspase.
Caspase 1 has also been found to play a role in processing a wide variety of proteins, most notably several cytokines and enzymes within the glycolytic pathway. The creation of the inflammasome during host responses to pathogens leads to the activation of caspase 1 and may induce the class of cell death known as pyroptosis. Caspase 1-mediated cleavage of the pro-inflammatory cytokine interleukin 1 beta (IL-1β) results in the biologically active form of this critical immune response regulator. Furthermore, inflammation-related disease models have illustrated a role for caspase 1 in asthma, rheumatoid arthritis, multiple sclerosis and other disorders.
FLICA (Fluorescent Labeled Inhibitor of Caspases) probes are comprised of an inhibitor peptide sequence that binds to active caspase enzymes, a fluoromethyl ketone (FMK) moiety that results in an irreversible binding event with the enzyme, and a fluorescent tag reporter. For a caspase 1 inhibitor, the multi-enzyme recognition sequence is tyrosine-valine-alanine-aspartic acid (YVAD). The FLICA® FAM-YVAD-FMK caspase-1 probe interacts with the enzymatic reactive center of activated caspase 1 via the YVAD recognition sequence, forming a covalent thioether adduct with the enzyme through the FMK moiety.
FLICA™ is cell permeant and non-cytotoxic. Unbound FLICA™ FAM-YVAD-FMK reagent is easily washed away; the remaining green fluorescent signal is a direct measure of caspase 1 activity at the time the probe was added. Detection of nuclear morphology and necrosis is also possible with the additional kit components, Hoechst 33342 and Propidium Iodide, respectively.
