Human Caspase 8 PicoKine ELISA Kit | CASP8 elisa kit

Human Caspase 8 PicoKine ELISA Kit

Cat. Num. AAA177624

• Gene Names: CASP8; CAP4; MACH; MCH5; FLICE; ALPS2B; Casp-8
• Reactivity: Human
• Synonyms: Caspase 8 PicoKine; Human Caspase 8 PicoKine ELISA Kit; Caspase-8; CASP-8; CASP8; CASP 8; Caspase8; Caspase 8; MACH; MCH5; MCH 5; CAP 4; CAP4; CAP-4; FADD Like ICE; caspase 8; CASP8 elisa kit

• Reactivity: Human
• Specificity: Natural and recombinant human Caspase 8
• Sequence Length: 464

• Assay Range: 31.2pg/ml-2000pg/ml
• Sensitivity: <10pg/ml
• Sample: For quantitative detection of human Caspase 8 in cell culture supernates, tissue homogenates and serum.
• Immunogen: E Coli,S217-D384&L385-D479
• Assay Procedure Step 1: Aliquot 0.1ml per well of the 2000pg/ml,1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml human Caspase 8 standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates, tissue homogenates or serum to each empty well. It is recommended that each human Caspase 8 standard solution and each sample be measured in duplicate.
• Intra-Assay Precision: Inter-Assay Precision; Sample|1|2|3|1|2|3; n|16|16|16|24|24|24; Mean(pg/ml)|153|589|1527|170|836|1721; Standard deviation|6.43|28.86|80.93|11.05|59.36|141.12; CV(%)|4.2|4.9|5.3|6.5|7.1|8.2
• Sample Data: Concentration(pg/ml)|0|31.2|62.5|125|250|500|1000|2000; O.D.|0.003|0.067|0.111|0.218|0.407|0.820|1.339|2.191
• Preparation and Storage: Store at 4 degree C for 6 months, at -20 degree C for 12 months. Avoid multiple freeze-thaw cycles.

Typical Testing Data/Standard Curve (for reference only)

(Human Caspase 8 PicoKine ELISA Kit standard curve )

Related Product Information for CASP8 elisa kit

Principle of the assay: Human Caspase 8 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Caspase 8 has been precoated onto 96-well plates. Standards(Expression system for standard: E Coli, Immunogen sequence: S217-D384 & L385-D479) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Caspase 8 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Caspase 8 amount of sample captured in plate.

Background: Caspase-8 is a caspase protein, encoded by the CASP8 gene. It is mapped to human chromosome 2q33-q34 and mouse chromosome 1B-proximal C. This gene encodes 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. This protein is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with Fas-interacting protein FADD. And this protein was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases. Many alternatively spliced transcript variants encoding different isoforms have been described, although not all variants have had their full-length sequences determined.

NCBI and Uniprot Product Information

• NCBI GI #: 122056474
• NCBI GeneID: 841
• NCBI Accession #: NP_001073593.1
• NCBI GenBank Nucleotide #: NM_001080124.1
• UniProt Accession #: Q14790; O14676; Q14791; Q14792; Q14793; Q14794; Q14795; Q14796; Q15780; Q15806; Q53TT5; Q8TDI1
• Molecular Weight: 61,836 Da
• NCBI Official Full Name: caspase-8 isoform C
• NCBI Official Synonym Full Names: caspase 8
• NCBI Official Symbol: CASP8
• NCBI Official Synonym Symbols: CAP4; MACH; MCH5; FLICE; ALPS2B; Casp-8
• NCBI Protein Information: caspase-8
• UniProt Protein Name: Caspase-8
• Protein Family: Caspase
• UniProt Gene Name: CASP8
• UniProt Synonym Gene Names: MCH5; CASP-8; FLICE; MACH
• UniProt Entry Name: CASP8_HUMAN

NCBI Description

This gene encodes 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. This protein is involved in the programmed cell death induced by Fas and various apoptotic stimuli. The N-terminal FADD-like death effector domain of this protein suggests that it may interact with Fas-interacting protein FADD. This protein was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases. Many alternatively spliced transcript variants encoding different isoforms have been described, although not all variants have had their full-length sequences determined. [provided by RefSeq, Jul 2008]

Uniprot Description

CASP8: 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. 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. Interacts with human cytomegalovirus/HHV-5 protein vICA/UL36; this interaction inhibits CASP8 activation. 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. Belongs to the peptidase C14A family. 9 isoforms of the human protein are produced by alternative splicing.

Protein type: Apoptosis; EC 3.4.22.61; Protease

Chromosomal Location of Human Ortholog: 2q33-q34

Cellular Component: CD95 death-inducing signaling complex; cytoplasm; cytoskeleton; cytosol; lipid raft; microtubule organizing center; mitochondrial outer membrane; mitochondrion; neuron projection; nucleoplasm

Molecular Function: cysteine-type endopeptidase activity; cysteine-type peptidase activity; death receptor binding; peptidase activity; protein binding; protein complex binding; tumor necrosis factor receptor binding; ubiquitin protein ligase binding

Biological Process: apoptosis; B cell activation; caspase activation; cell structure disassembly during apoptosis; cell surface receptor linked signal transduction; macrophage differentiation; natural killer cell activation; negative regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of macrophage differentiation; positive regulation of proteolysis; protein heterooligomerization; proteolysis; proteolysis involved in cellular protein catabolic process; response to antibiotic; response to cobalt ion; response to cold; response to estradiol stimulus; response to ethanol; response to lipopolysaccharide; T cell activation

Disease: Breast Cancer; Caspase 8 Deficiency; Hepatocellular Carcinoma; Lung Cancer