Caspase 8 Antibody - Internal - AF6442

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$280 100ul
$350 200ul

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  • Product Name
    Caspase 8 Antibody
  • Catalog No.
    AF6442
  • RRID
    AB_2835266
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Pig(80%), Bovine(90%), Horse(80%), Sheep(80%), Rabbit(90%), Dog(90%)
  • UniProt
  • Mol.Wt
    (Observed)55kD; (Calculated)55kDa
  • Concentration
    1mg/ml

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 #AF6442-BP Caspase 8 Blocking Peptide $350/1mg

Product Information

Alternative Names:Expand▼

ALPS2B; Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein; Apoptotic cysteine protease; Apoptotic protease Mch-5; Apoptotic protease Mch5; CAP4; CASP-8; CASP8; CASP8_HUMAN; Caspase 8; Caspase 8 apoptosis related cysteine peptidase; Caspase-8 subunit p10; CED 3; FADD Like ICE; FADD-homologous ICE/CED-3-like protease; FADD-like ICE; FLICE; FLJ17672; ICE-like apoptotic protease 5; MACH alpha 1/2/3 protein; MACH; MACH beta 1/2/3/4 protein; MCH5; MGC78473; MORT1 associated ced 3 homolog; MORT1-associated CED-3 homolog; OTTHUMP00000163717; OTTHUMP00000163720; OTTHUMP00000163724; OTTHUMP00000163725; OTTHUMP00000165062; OTTHUMP00000165063; OTTHUMP00000165064; OTTHUMP00000206552; OTTHUMP00000206582;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(80%), Bovine(90%), Horse(80%), Sheep(80%), Rabbit(90%), Dog(90%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Specificity:

Caspase 8 Antibody detects endogenous levels of total Caspase 8.

RRID:

AB_2835266
Please cite this product as: Affinity Biosciences Cat# AF6442, RRID:AB_2835266.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

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.

Immunogen Information

Immunogen:

A synthesized peptide derived from human Caspase 8, corresponding to a region within the internal amino acids.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

CASP8

Molecular Weight:

Observed Mol.Wt.: (Observed)55kD.
Predicted Mol.Wt.: (Calculated)55kDa.

Subcellular Location:

Cytoplasm.

Tissue Specificity:

Q14790 CASP8_HUMAN:
Isoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle.

Description:

This gene encodes a protein that is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes composed of a prodomain, a large protease subunit, and a small protease subunit. Activation of caspases requires proteolytic processing at conserved internal aspartic residues to generate a heterodimeric enzyme consisting of the large and small subunits.

Sequence:
MDFSRNLYDIGEQLDSEDLASLKFLSLDYIPQRKQEPIKDALMLFQRLQEKRMLEESNLSFLKELLFRINRLDLLITYLNTRKEEMERELQTPGRAQISAYRVMLYQISEEVSRSELRSFKFLLQEEISKCKLDDDMNLLDIFIEMEKRVILGEGKLDILKRVCAQINKSLLKIINDYEEFSKERSSSLEGSPDEFSNGEELCGVMTISDSPREQDSESQTLDKVYQMKSKPRGYCLIINNHNFAKAREKVPKLHSIRDRNGTHLDAGALTTTFEELHFEIKPHDDCTVEQIYEILKIYQLMDHSNMDCFICCILSHGDKGIIYGTDGQEAPIYELTSQFTGLKCPSLAGKPKVFFIQACQGDNYQKGIPVETDSEEQPYLEMDLSSPQTRYIPDEADFLLGMATVNNCVSYRNPAEGTWYIQSLCQSLRERCPRGDDILTILTEVNYEVSNKDDKKNMGKQMPQPTFTLRKKLVFPSD

Research Background

Function:

Most upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp-|-AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex (PubMed:23516580, PubMed:9006941). Cleaves RIPK1 at 'Asp-325' which is crucial for limiting apoptosis and necroptosis during embryonic development (By similarity).

Post-translational Modifications:

Generation of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events.

Phosphorylation on Ser-387 during mitosis by CDK1 inhibits activation by proteolysis and prevents apoptosis. This phosphorylation occurs in cancer cell lines, as well as in primary breast tissues and lymphocytes.

Subcellular Location:

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Tissue Specificity:

Isoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle.

Subunit Structure:

Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 18 kDa (p18) and a 10 kDa (p10) subunit (PubMed:10508784). Interacts with FADD, CFLAR and PEA15 (PubMed:10442631). Isoform 9 interacts at the endoplasmic reticulum with a complex containing BCAP31, BAP29, BCL2 and/or BCL2L1 (PubMed:11917123, PubMed:9334338). Interacts with TNFAIP8L2 (By similarity). Interacts with CASP8AP2 (PubMed:16378960). Interacts with RFFL and RNF34; negatively regulate CASP8 through proteasomal degradation (PubMed:15069192). Interacts with NOL3; decreases CASP8 activity in a mitochondria localization- and phosphorylation-dependent manner and this interaction is dissociated by calcium (PubMed:15509781). Interacts with UBR2 (PubMed:28602583). Interacts with RIPK1 (By similarity). Interacts with stimulated TNFRSF10B; this interaction is followed by CASP8 proteolytic cleavage and activation (PubMed:18846110).

(Microbial infection) Interacts with human cytomegalovirus/HHV-5 protein vICA/UL36; this interaction inhibits CASP8 activation.

(Microbial infection) Interacts with NleF from pathogenic E.coli.

(Microbial infection) Interacts with molluscum contagiosum virus protein MC160.

Similarity:

Isoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex.

Belongs to the peptidase C14A family.

Research Fields

Research Fields:

· Cellular Processes > Cell growth and death > p53 signaling pathway.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis - multiple species.(View pathway)
· Cellular Processes > Cell growth and death > Necroptosis.(View pathway)
· Environmental Information Processing > Signal transduction > TNF signaling pathway.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cardiovascular diseases > Viral myocarditis.
· Human Diseases > Infectious diseases: Bacterial > Legionellosis.
· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.
· Human Diseases > Cancers: Overview > Viral carcinogenesis.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Neurodegenerative diseases > Huntington's disease.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > Toll-like receptor signaling pathway.(View pathway)
· Organismal Systems > Immune system > IL-17 signaling pathway.(View pathway)
· Organismal Systems > Immune system > NOD-like receptor signaling pathway.(View pathway)

Protocols & Troubleshooting

Reference Citations:

1). Liu Y et al. Increased autophagy in EOC re-ascites cells can inhibit cell death and promote drug resistance. Cell Death Dis 2018 Apr 1;9(4):419 (PubMed: 29549251) [IF=5.959]
2). Gu LL et al. Oxymatrine Causes Hepatotoxicity by Promoting the Phosphorylation of JNK and Induction of Endoplasmic Reticulum Stress Mediated by ROS in LO2 Cells. Mol Cells 2018 May 31;41(5):401-412 (PubMed: 29754474) [IF=3.533]
3). Bai Y et al. BCL2L10 inhibits growth and metastasis of hepatocellular carcinoma both in vitro and in vivo. Mol Carcinog 2017 Mar;56(3):1137-1149 (PubMed: 27770580) [IF=3.411]

Application: WB    Species:mouse;    Sample:Not available

Figure 3. BCL2L10 induced cell apoptosis and inhibited tumor growth in nude mice. (A) BCL2L10 induced apoptosis in HepG2 and Huh7 cells, as determined by flow cytometry analysis following Annexin V and PI staining. The upper panel represents FACS images of HCC cells transfected with empty vector or BCL2L10, while the lower panel shows the quantitative analyses of early apoptotic and late apoptotic cells. The experiment was repeated three times in triplicate. Data are mean ± SD. (B) Protein expression of the active forms of apoptosis related genes caspase 3, caspase 8, and caspase 9 was evaluated by Western blot.

4). Ren W et al. Arginine inhibits the malignant transformation induced by interferon-gamma through the NF-κB-GCN2/eIF2α signaling pathway in mammary epithelial cells in vitro and in vivo. Exp Cell Res 2018 Jul 15;368(2):236-247 (PubMed: 29746817) [IF=3.329]
5). Yi J et al. Protective mechanisms of purified polyphenols from pinecones of Pinus koraiensis on spleen tissues in tumor-bearing S180 mice in vivo. Food Funct 2017 Jan 25;8(1):151-166 (PubMed: 27924972) [IF=3.241]

Application: IHC    Species:mouse;    Sample:Not available

Fig. 15 Effect of PPP-40 on caspase-8 expression of splenocytes in S180 mice (magnification × 400). The arrows show the positive expression of caspase-8. (A) Normal group; (B) model group; (C) positive control group (CTX 20 mg kg−1 ); (D) low concentration of the PPP-40 group (50 mg kg−1 ); (E) medium concentration of the PPP-40 group (150 mg kg−1 ); (F) high concentration of the PPP-40 group (300 mg kg−1 ); (G) quantity analysis (mean ± SD, n = 6). *p < 0.05 and **p < 0.01 compared with the normal group; #p < 0.05 and ##p < 0.01 compared with the model group.

6). Shen Z et al. Osthole induced apoptosis in human normal liver cells by regulating cell proliferation and endoplasmic reticulum stress. Environ Toxicol 2019 Mar 8 (PubMed: 30848542)
7). Li Y et al. Apoptotic effects of rhein through the mitochondrial pathways, two death receptor pathways, and reducing autophagy in human liver L02 cells. Environ Toxicol 2019 Aug 21 (PubMed: 31436023)
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Catalog Number :

AF6442-BP
(Blocking peptide available as AF6442-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide（0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Rabbit
90%
Bovine
90%
Dog
90%
Horse
80%
Sheep
80%
Pig
80%
Chicken
70%
Xenopus
0%
Zebrafish
0%
High similarity Medium similarity Low similarity No similarity
Q14790 as Substrate
Site PTM Type Enzyme
S16 Phosphorylation
K23 Ubiquitination
K34 Ubiquitination
K39 Ubiquitination
K63 Ubiquitination
S113 Phosphorylation
K121 Ubiquitination
K130 Ubiquitination
K148 Ubiquitination
K156 Acetylation
K156 Ubiquitination
K161 Ubiquitination
K169 Ubiquitination
K173 Ubiquitination
Y178 Phosphorylation
K183 Ubiquitination
S192 Phosphorylation
S209 Phosphorylation
S211 Phosphorylation
S219 Phosphorylation
K224 Ubiquitination
K229 Ubiquitination
Y235 Phosphorylation
K253 Ubiquitination
T263 Phosphorylation P51812 (RPS6KA3)
T273 Phosphorylation
S305 Phosphorylation
Y334 Phosphorylation
K344 Ubiquitination
S347 Phosphorylation Q16539 (MAPK14)
K351 Ubiquitination
K353 Ubiquitination
T373 Phosphorylation
S375 Phosphorylation
Y380 Phosphorylation P07948 (LYN) , P12931 (SRC)
S387 Phosphorylation P28482 (MAPK1) , P06493 (CDK1) , P27361 (MAPK3)
Y448 Phosphorylation P07948 (LYN)
K461 Ubiquitination
K473 Ubiquitination
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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