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Caspase 4 Assay Kit (Fluorometric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEVD.

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Publications

Key facts

Detection method

Fluorescent

Sample types

Tissue Extracts, Cell Lysate

Assay type

Enzyme activity

Assay time

2h

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Target data

Function

Inflammatory caspase that acts as an essential effector of NLRP3 inflammasome-dependent CASP1 activation and IL1B and IL18 secretion in response to non-canonical activators, such as UVB radiation, cholera enterotoxin subunit B and cytosolic LPS (PubMed:23516580, PubMed:24879791, PubMed:25119034, PubMed:22246630, PubMed:26174085, PubMed:26173988, PubMed:26508369, PubMed:25964352). Thiol protease that cleaves a tetrapeptide after an Asp residue at position P1 (PubMed:7797510, PubMed:23516580). Independently of NLRP3 inflammasome and CASP1, promotes pyroptosis, through GSDMD cleavage and activation, followed by IL1A, IL18 and HMGB1 release in response to non-canonical inflammasome activators (PubMed:26375003, PubMed:32109412). Plays a crucial role in the restriction of Salmonella typhimurium replication in colonic epithelial cells during infection: in later stages of the infection, LPS from cytosolic Salmonella triggers CASP4 activation, which catalyzes cleavage of GSDMD, resulting in pyroptosis of infected cells and their extrusion into the gut lumen, as well as in IL18 secretion (PubMed:25121752, PubMed:26375003, PubMed:25964352, PubMed:32109412). Cleavage of GSDMD is not strictly dependent on the consensus cleavage site but depends on an exosite interface on CASP4 that recognizes and binds the Gasdermin-D, C-terminal (GSDMD-CT) part (PubMed:32109412). Pyroptosis limits bacterial replication, while cytokine secretion promotes the recruitment and activation of immune cells and triggers mucosal inflammation (PubMed:25121752, PubMed:26375003, PubMed:25964352). Involved in LPS-induced IL6 secretion; this activity may not require caspase enzymatic activity (PubMed:26508369). Involved in cell death induced by endoplasmic reticulum stress and by treatment with cytotoxic APP peptides found Alzheimer's patient brains (PubMed:15123740, PubMed:22246630, PubMed:23661706). Activated by direct binding to LPS without the need of an upstream sensor (PubMed:25119034). Does not directly process IL1B (PubMed:7743998, PubMed:7797510, PubMed:7797592). During non-canonical inflammasome activation, cuts CGAS and may play a role in the regulation of antiviral innate immune activation (PubMed:28314590).

Alternative names

What's included?

100 Test
Components
2X Reaction Buffer I
4 x 2 mL
DTT I
1 x 400 µL
LEVD-AFC
1 x 500 µL
Lysis Buffer IV
1 x 100 mL

Recommended products

Caspase 4 Assay Kit (Fluorometric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEVD.

Alternative names

Key facts

Detection method

Fluorescent

Sample types

Tissue Extracts, Cell Lysate

Assay type

Enzyme activity

Assay time

2h

Assay Platform

Microplate reader

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Abcam's Caspase 4 Assay Kit (Fluorometric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEVD. The assay is based on detection of cleavage of substrate LEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). LEVD-AFC emits blue light (λmax = 400 nm); upon cleavage of the substrate by caspase 4 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified using a fluorometer or a fluorecence microtiter plate reader. Visit our for tips and troubleshooting.

Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. **Other caspase and apoptosis assays** Review the full set of , or the .

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