Caspase 8 Antibody | Affinity Biosciences

Product:	Caspase 8 Antibody
Catalog:	AF6442
Description:	Rabbit polyclonal antibody to Caspase 8
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat, Monkey
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog
Mol.Wt.:	55kDa; 55kD(Calculated).
Uniprot:	Q14790
RRID:	AB_2835266

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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. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

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Related Downloads

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, IF/ICC 1:100-1:500
*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,Monkey

Prediction:

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

Clonality:

Polyclonal

Specificity:

Caspase 8 Antibody detects endogenous levels of total Caspase 8.

RRID:

AB_2835266
Cite Format: Affinity Biosciences Cat# AF6442, RRID:AB_2835266.

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

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;

Immunogens

Immunogen:

Uniprot:

Q14790(CASP8_HUMAN) >>Visit HPA database.

Gene(ID):

CASP8(841)

Expression:

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:

Q14790 CASP8_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MDFSRNLYDIGEQLDSEDLASLKFLSLDYIPQRKQEPIKDALMLFQRLQEKRMLEESNLSFLKELLFRINRLDLLITYLNTRKEEMERELQTPGRAQISAYRVMLYQISEEVSRSELRSFKFLLQEEISKCKLDDDMNLLDIFIEMEKRVILGEGKLDILKRVCAQINKSLLKIINDYEEFSKERSSSLEGSPDEFSNGEELCGVMTISDSPREQDSESQTLDKVYQMKSKPRGYCLIINNHNFAKAREKVPKLHSIRDRNGTHLDAGALTTTFEELHFEIKPHDDCTVEQIYEILKIYQLMDHSNMDCFICCILSHGDKGIIYGTDGQEAPIYELTSQFTGLKCPSLAGKPKVFFIQACQGDNYQKGIPVETDSEEQPYLEMDLSSPQTRYIPDEADFLLGMATVNNCVSYRNPAEGTWYIQSLCQSLRERCPRGDDILTILTEVNYEVSNKDDKKNMGKQMPQPTFTLRKKLVFPSD

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

Dog

Rabbit

Pig

Horse

Sheep

Chicken

Xenopus

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q14790 As Substrate

Q14790

Site	PTM Type	Enzyme	Source
S16	Phosphorylation		Uniprot
K23	Ubiquitination		Uniprot
K34	Ubiquitination		Uniprot
K39	Ubiquitination		Uniprot
K63	Ubiquitination		Uniprot
S113	Phosphorylation		Uniprot
K121	Ubiquitination		Uniprot
K130	Ubiquitination		Uniprot
K148	Ubiquitination		Uniprot
K156	Acetylation		Uniprot
K156	Ubiquitination		Uniprot
K161	Ubiquitination		Uniprot
K169	Ubiquitination		Uniprot
K173	Ubiquitination		Uniprot
Y178	Phosphorylation		Uniprot
K183	Ubiquitination		Uniprot
S192	Phosphorylation		Uniprot
S209	Phosphorylation		Uniprot
S211	Phosphorylation		Uniprot
S219	Phosphorylation		Uniprot
K224	Ubiquitination		Uniprot
K229	Ubiquitination		Uniprot
Y235	Phosphorylation		Uniprot
K253	Ubiquitination		Uniprot
T263	Phosphorylation	P51812 (RPS6KA3)	Uniprot
T273	Phosphorylation		Uniprot
S305	Phosphorylation		Uniprot
Y334	Phosphorylation		Uniprot
K344	Ubiquitination		Uniprot
S347	Phosphorylation	Q16539 (MAPK14)	Uniprot
K351	Ubiquitination		Uniprot
K353	Ubiquitination		Uniprot
T373	Phosphorylation		Uniprot
S375	Phosphorylation		Uniprot
Y380	Phosphorylation	P07948 (LYN) , P12931 (SRC)	Uniprot
S387	Phosphorylation	P28482 (MAPK1) , P06493 (CDK1) , P27361 (MAPK3)	Uniprot
Y448	Phosphorylation	P07948 (LYN)	Uniprot
K461	Ubiquitination		Uniprot
K473	Ubiquitination		Uniprot

Research Backgrounds

Q14790_CASP8_HUMAN

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. Cleaves RIPK1 at 'Asp-325' which is crucial for limiting apoptosis and necroptosis during embryonic development (By similarity).

PTMs:

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.

Tissue Specificity:

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. Interacts with FADD, CFLAR and PEA15. Isoform 9 interacts at the endoplasmic reticulum with a complex containing BCAP31, BAP29, BCL2 and/or BCL2L1. Interacts with TNFAIP8L2 (By similarity). Interacts with CASP8AP2. Interacts with RFFL and RNF34; negatively regulate CASP8 through proteasomal degradation. Interacts with NOL3; decreases CASP8 activity in a mitochondria localization- and phosphorylation-dependent manner and this interaction is dissociated by calcium. Interacts with UBR2. Interacts with RIPK1 (By similarity). Interacts with stimulated TNFRSF10B; this interaction is followed by CASP8 proteolytic cleavage and activation.

(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.

Family&Domains:

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

Belongs to the peptidase C14A family.

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 > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Human Diseases > Neurodegenerative diseases > Huntington's disease.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cardiovascular diseases > Viral myocarditis.

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

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

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

· Organismal Systems > Immune system > IL-17 signaling pathway. (View pathway)

References

1). Increased autophagy in EOC re-ascites cells can inhibit cell death and promote drug resistance. Cell Death & Disease (PubMed: 29549251) [IF=9.0]

Application: IHC Species: human Sample: ascites cells

Fig. 5 |Apoptosis is increased in the EOC chemosensitive group. a, b IHC and IF analysis of apoptotic protein expression in ovarian cancer samples. c, d Western blot analysis of apoptotic proteins expressed in the no-chemotherapy group and chemosensitive group (***p<0.001,**p<0.01, *p<0.05). e qRT-PCR analysis of the average relative mRNA expression levels of apoptosis-related genes in the no-chemotherapy group and chemosensitive group (*p<0.05)

Application: IF/ICC Species: human Sample: ascites cells

Application: WB Species: human Sample: ascites cells

2). Silica nanoparticles cause spermatogenesis dysfunction in mice via inducing cell cycle arrest and apoptosis. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY (PubMed: 35051769) [IF=6.8]

Application: WB Species: mouse Sample: testis

Fig. 7. |SiNPs affected the protein expressions of ATM/p53 and TNF-α/TNFR I-mediated signaling pathways. (A) The protein expressions of ATM, p53, Bcl-2, Bax,TNF-α, TNFR I, Caspase-8, and Caspase-3 were detected in the testis of control group and SiNPs group.

3). Protective mechanisms of purified polyphenols from pinecones of Pinus koraiensis on spleen tissues in tumor-bearing S180 mice in vivo. Food & Function (PubMed: 27924972) [IF=6.1]

Application: IHC Species: mouse Sample:

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.

4). Naringin ameliorates type 2 diabetes mellitus-induced steatohepatitis by inhibiting RAGE/NF-κB mediated mitochondrial apoptosis. LIFE SCIENCES (PubMed: 32702445) [IF=6.1]

Application: WB Species: rat Sample: liver

Fig. 7. |Naringin abrogates mitochondrial apoptosis. (H–J) caspase 8, caspase 9, and caspase 3 were determined by immunoblotting in liver tissue. Mean ± S.E.M (n = 3), *Control vs NASH; #NASH vs NAR. *P < 0.05, **P < 0.01, ***P < 0.001; #P < 0.05, ##P < 0.01, ###P < 0.001.

5). Akkermansia muciniphila Aspartic Protease Amuc_1434* Inhibits Human Colorectal Cancer LS174T Cell Viability via TRAIL-Mediated Apoptosis Pathway. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (PubMed: 32403433) [IF=5.6]

Application: WB Species: human Sample: LS174T cells

Figure 5. | Amuc_1434* mediated the activation of the apoptosis pathway in LS174T cells. (A) The expression of death receptor 4 (DR4), death receptor 5 (DR5), cysteinyl aspartate specific proteinase 8(caspase 8) and cysteinyl aspartate specific proteinase 3 (caspase 3) induced by Amuc_1434* in LS174T cells was dependent on tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). (a) LS174T cells were treated with 8 and 64 µg/mL Amuc_1434* for 24 h, respectively. The cell lysates were analyzed by Western blot.

6). SOX11 and FAK participate in the stretch‑induced mechanical injury to alveolar type 2 epithelial cells. International Journal of Molecular Medicine (PubMed: 33236128) [IF=5.4]

Application: WB Species: Human Sample: AT2 cells

Figure 6 (A-C) Western blot analysis showing caspase-3/8 expression in AT2 cells. The overexpression of SOX11 inhibited, whilst the knockdown of SOX11 increased caspase-3/8 expression. Furthermore, FAK antagonism blocked the effect of SOX11 overexpression on caspase-3/8 expression. Data are presented as the means ± standard error of the mean. *P<0.05, **P<0.01. AT2, alveolar type II; SOX, Sex-determining gene on the Y chromosome related high mobility group box; FAK, focal adhesion kinase.

7). Bisimidazolium Salt Glycosyltransferase Inhibitors Suppress Hepatocellular Carcinoma Progression In Vitro and In Vivo. Pharmaceuticals (PubMed: 35745636) [IF=4.6]

Application: WB Species: Human Sample: HepG2 cells

Figure 8 Treatment with C20/C22 led to increased proapoptotic protein expression and a caspase cascade response. (A–C) WB analysis of the antiapoptotic and caspase proteins in HepG2 cells treated with C20/C22 at 2 μM for 0, 6, 12, and 24 h. The reported values correspond to the mean ± SD for three independent experiments. (D) Immunofluorescence assays employed to determine the expression of caspase3/7 in cells with indicated treatments. Scale bars, 10 µm. (E,F) Apoptotic cells detected by flow cytometry after indicated treatments (n = 3). The p-value was analyzed by one-way ANOVA followed by Tukey’s test using GraphPad Prism version 8.00. ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. 0 h/control group.

8). BCL2L10 inhibits growth and metastasis of hepatocellular carcinoma both in vitro and in vivo. MOLECULAR CARCINOGENESIS (PubMed: 27770580) [IF=4.6]

Application: WB Species: mouse Sample:

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.

9). Osthole induced apoptosis in human normal liver cells by regulating cell proliferation and endoplasmic reticulum stress. ENVIRONMENTAL TOXICOLOGY (PubMed: 30848542) [IF=4.5]

10). Apoptotic effects of rhein through the mitochondrial pathways, two death receptor pathways, and reducing autophagy in human liver L02 cells. ENVIRONMENTAL TOXICOLOGY (PubMed: 31436023) [IF=4.5]

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Caspase 8 Antibody - #AF6442