Caspase-1 Assay Kit (Fluorometric)
Detection method: Fluorescent
Example type: Tissue extracts, cell lysate
Test type: Enzymatic activity
Rehearsal time: 2h00m
Caspase-1 Assay Kit (Fluorometric) (ab39412) provides a simple and convenient method to detect caspase-1 activity, which recognizes the YVAD sequence. The caspase-1 assay protocol is based on the cleavage of the substrate YVAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). YVAD-AFC emits blue light (Em=400 nm); upon cleavage of the substrate by caspase-1 or related caspases, free AFC fluoresces yellow-green (Ex/Em=400/505 nm), which can be quantified using a fluorometer or fluorescence microtiter plate reader. Comparison of the fluorescence of a treated sample with an untreated control allows the determination of the fold increase in caspase-1 activity.
- Caspase-1 Assay Protocol Summary:
- add samples to wells
- add reaction buffer and YVAD-AFC substrate and incubate for 1-2 hours
- analyze with a microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called the K110 Caspase-1 Fluorometric Assay Kit. K110-100 is the same size as the 100 test size of ab39412. Caspase-1 (ICE, IL-1beta Converting Enzyme) is the prototypical member of the ICE family of proteases/caspases.
Platform: microplate reader
Store at -20°C. Consult the protocols.
Caspases are a family of cysteine proteases that are key mediators of programmed cell death, or apoptosis. The precursor form of all caspases is composed of a prodomain and large and small catalytic subunits. Active forms of caspases are generated by various stimuli, including ligand-receptor interactions, growth factor deprivation, and inhibitors of cellular functions.
All known caspases require cleavage adjacent to aspartates to release a large and a small subunit, which associates with an α2b2 tetramer to form the active enzyme. Caspase 1 is similar to the C. elegans cell death gene CED3 and regulates multiple proinflammatory cytokines, including interleukin 1b and interferon-inducing factor gamma. Caspase 1 plays a role downstream of Caspase 8 which is involved in Fas-mediated apoptosis.
Cell localization: cytoplasmic
- CASP1 nirs variant 1