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AB65658

Caspase 4 Assay Kit (Fluorometric)

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(2 Publications)

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

View Alternative Names

ICH2, CASP4, Caspase-4, CASP-4, ICE and Ced-3 homolog 2, ICE(rel)-II, Mih1, Protease TX, ICH-2

1 Images
Schematic Diagram - Caspase 4 Assay Kit (Fluorometric) (AB65658)
  • Schematic Diagram

Supplier Data

Schematic Diagram - Caspase 4 Assay Kit (Fluorometric) (AB65658)

Representative image of Caspase 4 Assay Kit (Fluorometric) ab65658

Components shown from left to right :

- LEVD-AFC

- DTT I

- 2X Reaction Buffer I (4 vials)

- Lysis Buffer IV

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.

Key facts

Detection method

Fluorescent

Sample types

Tissue Extracts, Cell Lysate

Assay type

Enzyme activity

Assay time

2h

Assay Platform

Microplate reader

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Enzyme activity assay": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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 .

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

What's included?

{ "values": { "100Test": { "sellingSize": "100 Test", "publicAssetCode":"ab65658-100Test", "assetComponentDetails": [ { "size":"1 x 100 mL", "name":"Lysis Buffer IV", "number":"AB65658-CMP04", "productcode":"AB323966" }, { "size":"1 x 0.5 mL", "name":"LEVD-AFC", "number":"AB65658-CMP03", "productcode":"" }, { "size":"1 x 400 µL", "name":"DTT I (1M)", "number":"AB65658-CMP02", "productcode":"" }, { "size":"4 x 2 mL", "name":"2X Reaction Buffer I", "number":"AB65658-CMP01", "productcode":"AB323963" } ] } } }

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Product protocols

Target data

Inflammatory caspase that acts as the effector of the non-canonical inflammasome by mediating lipopolysaccharide (LPS)-induced pyroptosis (PubMed : 25119034, PubMed : 26375003, PubMed : 32109412, PubMed : 34671164, PubMed : 37001519, PubMed : 37993712, PubMed : 37993714). Also indirectly activates the NLRP3 and NLRP6 inflammasomes (PubMed : 23516580, PubMed : 26375003, PubMed : 32109412, PubMed : 7797510). Acts as a thiol protease that cleaves a tetrapeptide after an Asp residue at position P1 : catalyzes cleavage of CGAS, GSDMD and IL18 (PubMed : 15326478, PubMed : 23516580, PubMed : 26375003, PubMed : 28314590, PubMed : 32109412, PubMed : 37993712, PubMed : 37993714, PubMed : 7797510). Effector of the non-canonical inflammasome independently of NLRP3 inflammasome and CASP1 : the non-canonical inflammasome promotes pyroptosis through GSDMD cleavage without involving secretion of cytokine IL1B (PubMed : 25119034, PubMed : 25121752, PubMed : 26375003, PubMed : 31268602, PubMed : 32109412, PubMed : 37993712, PubMed : 37993714). In the non-canonical inflammasome, CASP4 is activated by direct binding to the lipid A moiety of LPS without the need of an upstream sensor (PubMed : 25119034, PubMed : 25121752, PubMed : 29520027, PubMed : 32510692, PubMed : 32581219, PubMed : 37993712). LPS-binding promotes CASP4 activation and CASP4-mediated cleavage of GSDMD and IL18, followed by IL18 secretion through the GSDMD pore, pyroptosis of infected cells and their extrusion into the gut lumen (PubMed : 25119034, PubMed : 25121752, PubMed : 37993712, PubMed : 37993714). Also indirectly promotes secretion of mature cytokines (IL1A and HMGB1) downstream of GSDMD-mediated pyroptosis via activation of the NLRP3 and NLRP6 inflammasomes (PubMed : 26375003, PubMed : 32109412). Involved in NLRP3-dependent CASP1 activation and IL1B secretion in response to non-canonical activators, such as UVB radiation or cholera enterotoxin (PubMed : 22246630, PubMed : 23516580, PubMed : 24879791, PubMed : 25964352, PubMed : 26173988, PubMed : 26174085, PubMed : 26508369). Involved in NLRP6 inflammasome-dependent activation in response to lipoteichoic acid (LTA), a cell-wall component of Gram-positive bacteria, which leads to CASP1 activation and IL1B secretion (PubMed : 33377178). Involved in LPS-induced IL6 secretion; this activity may not require caspase enzymatic activity (PubMed : 26508369). The non-canonical inflammasome is required for innate immunity to cytosolic, but not vacuolar, bacteria (By similarity). Plays a crucial role in the restriction of S.typhimurium replication in colonic epithelial cells during infection (PubMed : 25121752, PubMed : 25964352). Activation of the non-canonical inflammasome in brain endothelial cells can lead to excessive pyroptosis, leading to blood-brain barrier breakdown (By similarity). Pyroptosis limits bacterial replication, while cytokine secretion promotes the recruitment and activation of immune cells and triggers mucosal inflammation (PubMed : 25121752, PubMed : 25964352, PubMed : 26375003). May also act as an activator of adaptive immunity in dendritic cells, following activation by oxidized phospholipid 1-palmitoyl-2-arachidonoyl- sn-glycero-3-phosphorylcholine, an oxidized phospholipid (oxPAPC) (By similarity). Involved in cell death induced by endoplasmic reticulum stress and by treatment with cytotoxic APP peptides found in Alzheimer's patient brains (PubMed : 15123740, PubMed : 22246630, PubMed : 23661706). 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). Catalyzes cleavage and maturation of IL18; IL18 processing also depends of the exosite interface on CASP4 (PubMed : 15326478, PubMed : 37993712, PubMed : 37993714). In contrast, it 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).. (Microbial infection) In response to the Td92 surface protein of the periodontal pathogen T.denticola, activated by cathepsin CTSG which leads to production and secretion of IL1A and pyroptosis of gingival fibroblasts.
See full target information CASP4

Publications (2)

Recent publications for all applications. Explore the full list and refine your search

Cellular & molecular biology letters 28:66 PubMed37587410

2023

MiR-15b-5p and PCSK9 inhibition reduces lipopolysaccharide-induced endothelial dysfunction by targeting SIRT4.

Applications

Unspecified application

Species

Unspecified reactive species

Elisa Martino,Nunzia D'Onofrio,Anna Balestrieri,Luigi Mele,Celestino Sardu,Raffaele Marfella,Giuseppe Campanile,Maria Luisa Balestrieri

Cell death and differentiation 29:1267-1282 PubMed34916628

2021

Cytoplasmic innate immune sensing by the caspase-4 non-canonical inflammasome promotes cellular senescence.

Applications

Unspecified application

Species

Unspecified reactive species

Irene Fernández-Duran,Andrea Quintanilla,Núria Tarrats,Jodie Birch,Priya Hari,Fraser R Millar,Anthony B Lagnado,Vanessa Smer-Barreto,Morwenna Muir,Valerie G Brunton,João F Passos,Juan Carlos Acosta
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