Human Caspase 4 (CASP4) ELISA Kit
Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
Principle of the Assay
The microtiter plate provided in this kit has been pre-coated with an antibody specific to CASP4. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to CASP4. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain CASP4, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of CASP4 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
For Use with serum, plasma, and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Target Information
Inflammatory caspase that acts as an essential effector of the NLRP3 and NLRP6 inflammasomes by mediating lipopolysaccharide (LPS)-induced pyroptosis (PubMed:7797510, PubMed:23516580, PubMed:26375003, PubMed:32109412). Thiol protease that cleaves a tetrapeptide after an Asp residue at position P1: catalyzes cleavage of CGAS, GSDMD and IL18 (PubMed:7797510, PubMed:15326478, PubMed:23516580, PubMed:26375003, PubMed:28314590, PubMed:32109412). Required for innate immunity to cytosolic, but not vacuolar, bacteria (By similarity). Plays a key role in NLRP3-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:26508369, PubMed:22246630, PubMed:23516580, PubMed:24879791, PubMed:25119034, PubMed:26173988, PubMed:26174085, PubMed:25964352). Activated by direct binding to LPS without the need of an upstream sensor (PubMed:25119034). 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 and IL18 secretion (PubMed:33377178). 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:34671164, 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 in Alzheimer's patient brains (PubMed:23661706, PubMed:15123740, PubMed:22246630). Catalyzes cleavage and maturation of IL18 (PubMed:15326478). In contrast, it does not directly process IL1B (PubMed:7743998, PubMed:7797592, PubMed:7797510). During non-canonical inflammasome activation, cuts CGAS and may play a role in the regulation of antiviral innate immune activation (PubMed:28314590).
GENE ID | 837 |
SWISS PROT | P49662 |
SYNONYMS |
ICE(rel)II; ICH2; TX; Protease ICH-2; Cysteinyl Aspartate Specific Proteinases 4; Apoptosis-Related Cysteine Peptidase |
Materials Supplied
Kit Components | 96 Wells Quantity/Size |
---|---|
Pre-coated, ready-to-use 96-well strip plate | 1 plate |
Plate sealer for 96 wells | 2 |
Standard |
2 tubes |
Diluent buffer | 1 bottle |
Detection Reagent A | 1 bottle |
Detection Reagent B | 1 bottle |
TMB Substrate | 1 tube |
Stop Solution | 1 tube |
Wash Buffer (30 ℅ concentrate) | 1 tube |
Product data sheet | 1 copy |
Storage
Storage | The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C. |
Performance Characteristics
REPEATABILITY |
Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level CASP4 were tested 20 times on one plate, respectively. |
SENSITIVITY | The minimum detectable dose was 0.062ng/mL. |
ASSAY RANGE | 0.156-10ng/mL |
SPECIFICITY | This assay has high sensitivity and excellent specificity for the detection of CASP4. No significant cross-reactivity or interference between CASP4 and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between CASP4 and all analogs, therefore, cross reactivity may still exist. |