Caspase 4 Antibody - #AF5130

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Product: Caspase 4 Antibody
Catalog: AF5130
Description: Rabbit polyclonal antibody to Caspase 4
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Bovine, Dog
Mol.Wt.: 35~50 kD; 43kD(Calculated).
Uniprot: P49662
RRID: AB_2837616

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 100ul $280 In stock
 200ul $350 In stock

Lead Time: Same day delivery

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Related Downloads
MS Report
HPLC Report
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,Mouse,Rat
Prediction:
Bovine(82%), Dog(82%)
Clonality:
Polyclonal
Specificity:
Caspase 4 Antibody detects endogenous levels of total Caspase 4.
RRID:
AB_2837616
Cite Format: Affinity Biosciences Cat# AF5130, RRID:AB_2837616.
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; CASP11;caspase 11;

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:
Strict requirement for Asp at the P1 position. It has a preferred cleavage sequence of Tyr-Val-Ala-Asp-|- but also cleaves at Asp-Glu-Val-Asp-|-. Ref.14 Enzyme regulation Inhibited by the effector protein NleF that is produced by pathogenic E.coli; this inhibits apoptosis.
Sequence:
MAEGNHRKKPLKVLESLGKDFLTGVLDNLVEQNVLNWKEEEKKKYYDAKTEDKVRVMADSMQEKQRMAGQMLLQTFFNIDQISPNKKAHPNMEAGPPESGESTDALKLCPHEEFLRLCKERAEEIYPIKERNNRTRLALIICNTEFDHLPPRNGADFDITGMKELLEGLDYSVDVEENLTARDMESALRAFATRPEHKSSDSTFLVLMSHGILEGICGTVHDEKKPDVLLYDTIFQIFNNRNCLSLKDKPKVIIVQACRGANRGELWVRDSPASLEVASSQSSENLEEDAVYKTHVEKDFIAFCSSTPHNVSWRDSTMGSIFITQLITCFQKYSWCCHLEEVFRKVQQSFETPRAKAQMPTIERLSMTRYFYLFPGN

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Bovine
82
Dog
82
Rabbit
64
Pig
0
Horse
0
Sheep
0
Xenopus
0
Zebrafish
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

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). Knockdown of NADPH oxidase 4 reduces mitochondrial oxidative stress and neuronal pyroptosis following intracerebral hemorrhage. Neural Regeneration Research (PubMed: 36751799) [IF=6.1]

Application: WB    Species: Rat    Sample:

Figure 4 Knockdown of NOX4 ameliorates neuronal pyroptosis through caspase 1/GSDMD-N and caspase4/11/GSDMD-C pathways after ICH. (A) Western blot analysis of Procaspase4/11, Cleaved caspase4/11 and Procaspase1 expression in the indicated groups. (B) Western blot analysis of GSDMD-FL, GSDMD-N and GSDMD-C expression in the indicated groups. (C–H) Quantitative analysis of Procaspase4/11 (C), Cleaved caspase4/11 (D), Procaspase1 (E), GSDMD-FL (F), GSDMD-N (G) and GSDMD-C (H) protein levels shown in A and B (n = 4 per group). (I) Representative immunofluorescence images showing the immunoreactivity of GSDMD (red) in neurons (NeuN-positive cells, yellow) in the indicated groups (AAV, GFP, green). The expression of GSDMD in the ICH and ICH + AAV-CON groups were increased, while that in the ICH + AAV-NOX4 group was decreased. Scale bars: 20 µm. (J) Representative immunohistochemical images illustrate the changes in the GSDMD protein expression in the indicated groups. The expression of GSDMD in the ICH and ICH + AAV-CON groups was increased, while that in the ICH + AAV-NOX4 group was decreased. Scale bars: 50 µm. (K, L) Percentage of GSDMD-positive cells in immunofluorescence (K) and immunohistochemical (L) images (n = 5 per group). Data are expressed as the mean ± SEM. ***P < 0.001 (one-way analysis of variance with Dunnett’s multiple comparison test). AAV-CON: AAV9-U6-shRNA (Scramble)-CMV-GFP; AAV-NOX4: AAV9-U6-shRNA (NOX4)-CMV-GFP; GSDMD: gasdermin D; ICH: intracerebral hemorrhage; MRI: magnetic resonance imaging; NOX4: nicotinamide adenine dinucleotide phosphate oxidase 4.
2). The protective effect of isoflurane pretreatment on liver IRI by suppressing noncanonical pyroptosis of liver macrophages. International Immunopharmacology (PubMed: 34332342) [IF=5.6]

Application: WB    Species: Mice    Sample: liver tissues

Fig. 2. Isoflurance pretreatement can attenuate liver injury by reducing noncanonical pyroptosis in liver macrophage A Caspase-11 expression in liver tissues was detected by Western biotting. B. Serum levels of IL-1β and IL-18 were measured. All data are shown as the mean ± SD n (sham) = 5 mice per group, n(IRI) = 5 mice per group *indicates statistical difference at P < 0.05.

Application: IF/ICC    Species: Mice    Sample: liver tissues

Fig. 3. Isoflurance pretreatement can attenuate liver injury by reducing noncanonical pyroptosis in liver macrophage A Representative fluorescence image of caspase-11 expression from liver sections(200x). B. Serum levels of IL-1β and IL-18 were measured. All data are shown as the mean ± SD n (sham) = 5 mice per group, n(IRI) = 5 mice per group *indicates statistical difference at P < 0.05.
3). Obeticholic acid improved triptolide/lipopolysaccharide-induced hepatotoxicity by inhibiting caspase-11-GSDMD pyroptosis pathway. Journal of Applied Toxicology [IF=3.3]
4). Ginsenoside Rg1 alleviates lipopolysaccharide‑induced neuronal damage by inhibiting NLRP1 inflammasomes in HT22 cells. Experimental and Therapeutic Medicine (PubMed: 34055081) [IF=2.7]
5). Inhibition of Caspase-11-Mediated Pyroptosis Alleviates Acute Kidney Injury Associated with Severe Acute Pancreatitis in Rats. JOURNAL OF INVESTIGATIVE SURGERY (PubMed: 36350036) [IF=1.9]

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