FAM-FLICA Caspase-1 Assay Kit

FAM-FLICA Caspase-1 Assay Kit

Product name

Caspase-1 Assay Kit (Fluorometric)

Detection method: Fluorescent

Example type: Tissue extracts, cell lysate

Test type: Enzymatic activity

Rehearsal time: 2h00m

Product Description

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

Notes

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

Properties

Storage instructions

Store at -20°C. Consult the protocols.

Relevance

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

Alternative names

  • CASP-1
  • CASP1
  • CASP1 nirs variant 1
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