Cleaved-Caspase 4 (Gln81) Antibody - #AF5373

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Product: Cleaved-Caspase 4 (Gln81) Antibody
Catalog: AF5373
Description: Rabbit polyclonal antibody to Cleaved-Caspase 4 (Gln81)
Application: WB IHC
Reactivity: Human
Mol.Wt.: 43kDa; 43kD(Calculated).
Uniprot: P49662
RRID: AB_2837858

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MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human
Clonality:
Polyclonal
Specificity:
Cleaved-Caspase4 (Gln81) Antibody detects endogenous levels of fragment of activated Caspase4 resulting from cleavage adjacent to Gln81.
RRID:
AB_2837858
Cite Format: Affinity Biosciences Cat# AF5373, RRID:AB_2837858.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Apoptotic cysteine protease Mih1/TX; CASP-4; CASP4; CASP4_HUMAN; Caspase 4 apoptosis related cysteine peptidase; Caspase-4 subunit 2; Caspase4; ICE(rel)-II; ICE(rel)II; ICEREL II; ICH2; Mih1/TX; Protease ICH-2; Protease TX; TX;

Immunogens

Immunogen:
Uniprot:

P49662(CASP4_HUMAN) >>Visit HPA database.

Gene(ID):
CASP4(837)
Expression:
P49662 CASP4_HUMAN:

Widely expressed, including in keratinocytes and colonic and small intestinal epithelial cells (at protein level). Not detected in brain.

Description:
Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves caspase-1.
Sequence:
MAEGNHRKKPLKVLESLGKDFLTGVLDNLVEQNVLNWKEEEKKKYYDAKTEDKVRVMADSMQEKQRMAGQMLLQTFFNIDQISPNKKAHPNMEAGPPESGESTDALKLCPHEEFLRLCKERAEEIYPIKERNNRTRLALIICNTEFDHLPPRNGADFDITGMKELLEGLDYSVDVEENLTARDMESALRAFATRPEHKSSDSTFLVLMSHGILEGICGTVHDEKKPDVLLYDTIFQIFNNRNCLSLKDKPKVIIVQACRGANRGELWVRDSPASLEVASSQSSENLEEDAVYKTHVEKDFIAFCSSTPHNVSWRDSTMGSIFITQLITCFQKYSWCCHLEEVFRKVQQSFETPRAKAQMPTIERLSMTRYFYLFPGN

PTMs - P49662 As Substrate

Site PTM Type Enzyme
S16 Phosphorylation
K49 Methylation
K53 Methylation
S83 Phosphorylation
K87 Ubiquitination
K107 Ubiquitination
K129 Ubiquitination
K225 Ubiquitination
S271 Phosphorylation
S274 Phosphorylation
T352 Phosphorylation

Research Backgrounds

Function:

Inflammatory caspase. 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. Independently of NLRP3 inflammasome and CASP1, promotes pyroptosis, through GSDMD cleavage and activation, and IL1A, IL18 and HMGB1 release in response to non-canonical inflammasome activators. 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 ultimately results in pyroptosis of infected cells and their extrusion into the gut lumen, as well as in IL18 secretion. Pyroptosis limits bacterial replication, while cytokine secretion promotes the recruitment and activation of immune cells and triggers mucosal inflammation. Involved in LPS-induced IL6 secretion; this activity may not require caspase enzymatic activity. Involved in cell death induced by endoplasmic reticulum stress and by treatment with cytotoxic APP peptides found Alzheimer's patient brains. Activated by direct binding to LPS without the need of an upstream sensor. Does not directly process IL1B. During non-canonical inflammasome activation, cuts CGAS and may play a role in the regulation of antiviral innate immune activation.

PTMs:

In response to activation signals, including endoplasmic reticulum stress or treatment with amyloid-beta A4 protein fragments (such as amyloid-beta protein 40), undergoes autoproteolytic cleavage.

Subcellular Location:

Cytoplasm>Cytosol. Endoplasmic reticulum membrane>Peripheral membrane protein>Cytoplasmic side. Mitochondrion. Inflammasome. Secreted.
Note: Predominantly localizes to the endoplasmic reticulum (ER). Association with the ER membrane requires TMEM214 (PubMed:15123740). Released in the extracellular milieu by keratinocytes following UVB irradiation (PubMed:22246630).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Widely expressed, including in keratinocytes and colonic and small intestinal epithelial cells (at protein level). Not detected in brain.

Subunit Structure:

Upon direct LPS-binding, forms large homooligomers, resulting in its activation. These oligomers are often referred to as 'non-canonical inflammasomes'. Active as a heterotetramer consisting of two anti-parallel arranged heterodimers, each one formed by a small and a large subunit (By similarity). In its precursor form, interacts with TMEM214; this interaction is required for association with the endoplasmic reticulum membrane. Interacts with CASP1. Interacts with NOD2. Interacts with SERPINB1; this interaction regulates CASP4 activity.

(Microbial infection) Interacts with NleF protein from pathogenic E.coli; this interaction leads to enzyme inhibition.

Family&Domains:

The CARD domain mediates LPS recognition and homooligomerization.

Belongs to the peptidase C14A family.

Research Fields

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

References

1). Inhibition of IL1R1 or CASP4 attenuates spinal cord injury through ameliorating NLRP3 inflammasome-induced pyroptosis. Frontiers in Immunology, 2022 (PubMed: 35990672) [IF=7.3]
2). Anoikis regulator GLI2 promotes NC cell immunity escape by TGF-β-mediated non-classic hedgehog signaling in colorectal cancer: based on artificial intelligence and big data analysis. Aging, 2023 (PubMed: 38159250) [IF=5.2]
3). Ochratoxin A induces cytotoxicity through ROS-mediated endoplasmic reticulum stress pathway in human gastric epithelium cells. TOXICOLOGY, 2022 (PubMed: 36058351) [IF=4.5]
4). Melatonin alleviates deoxynivalenol-induced apoptosis of human granulosa cells by reducing mutually accentuated FOXO1 and ER stress. Biology of Reproduction, 2021 (PubMed: 33907797) [IF=3.6]

Application: WB    Species: Human    Sample: granulosa cells

Figure 4. Melatonin attenuates DON-induced apoptosis by suppressing ER stress in human granulosa cells. (A) Cells were treated with different concentrations of DON for 24 h. The protein levels of BiP, CHOP, and phosphorylated-eIF2α were assessed by western blot. (B) For the inhibition of ROS production, Mito-TEMPO was added 1 h before DON treatment. Cells were cultured for another 24 h and the levels of BiP, CHOP, and phosphorylated-eIF2α were measured by western blot. (C) Granulosa cells were cultured in medium containing DON with or without melatonin for 24 h. The levels of BiP, CHOP, and phosphorylated-eIF2α were analyzed by western blot. (D) The levels of Cleaved Caspase 4 and Phosphorylated JNK were analyzed by western blot. (E) ER stress inhibitor 4-PBA was added 1 h before DON and melatonin incubation. The cells were cultured for 24 h and cell viability was determined using MTT assay kit. (F) The protein levels of Bax, Bcl-2, and Cleaved Caspase-3 from granulosa cells treated as depicted in Figure 4D were analyzed by western blot. (G) Quantification of Bcl-2 and Bax expression using densitometric analysis. β-actin served as the control for loading. Data are represented as means ± SEM of three independent experiments. ∗P < 0.05, ∗∗P < 0.01.

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