| Applications: | ELISA |
| Reactivity: | Human |
| Note: | FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: | 9.38pg/mL |
| Detection Limit: | 15.63~1000pg/mL |
| Short Description: | This A Beta 1-40 Sandwich ELISA is an in-vitro enzyme-linked immunosorbent assay for the measurement of samples in human cell culture supernatant, serum and plasma (EDTA, citrate, heparin). |
| Storage Instruction: | If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: | 3.5h |
| Detection: | Colormetric |
| Gene Symbol: | APP |
| Gene ID: | 351 |
| Uniprot ID: | A4_HUMAN |
| Specificity: | This kit recognizes Human A Beta 1-40 in samples. No significant cross-reactivity or interference between Human A Beta 1-40 and analogues was observed. |
| Sample Type: | Serum, plasma and other biological fluids |
| Post Translational Modifications | Proteolytically processed under normal cellular conditions. Cleavage either by alpha-secretase, beta-secretase or theta-secretase leads to generation and extracellular release of soluble APP peptides, S-APP-alpha and S-APP-beta, and the retention of corresponding membrane-anchored C-terminal fragments, C80, C83 and C99. Subsequent processing of C80 and C83 by gamma-secretase yields P3 peptides. This is the major secretory pathway and is non-amyloidogenic. Alternatively, presenilin/nicastrin-mediated gamma-secretase processing of C99 releases the amyloid-beta proteins, amyloid-beta protein 40 and amyloid-beta protein 42, major components of amyloid plaques, and the cytotoxic C-terminal fragments, gamma-CTF(50), gamma-CTF(57) and gamma-CTF(59). PSEN1 cleavage is more efficient with C83 than with C99 as substrate (in vitro). Amyloid-beta protein 40 and Amyloid-beta protein 42 are cleaved by ACE. Many other minor amyloid-beta peptides, amyloid-beta 1-X peptides, are found in cerebral spinal fluid (CSF) including the amyloid-beta X-15 peptides, produced from the cleavage by alpha-secretase and all terminating at Gln-686. Proteolytically cleaved by caspases during neuronal apoptosis. Cleavage at Asp-739 by either CASP6, CASP8 or CASP9 results in the production of the neurotoxic C31 peptide and the increased production of amyloid-beta peptides. N-glycosylated. N- and O-glycosylated. O-glycosylation on Ser and Thr residues with core 1 or possibly core 8 glycans. Partial tyrosine glycosylation (Tyr-681) is found on some minor, short amyloid-beta peptides (amyloid-beta 1-15, 1-16, 1-17, 1-18, 1-19 and 1-20) but not found on amyloid-beta protein 38, amyloid-beta protein 40 nor on amyloid-beta protein 42. Modification on a tyrosine is unusual and is more prevelant in AD patients. Glycans had Neu5AcHex(Neu5Ac)HexNAc-O-Tyr, Neu5AcNeu5AcHex(Neu5Ac)HexNAc-O-Tyr and O-AcNeu5AcNeu5AcHex(Neu5Ac)HexNAc-O-Tyr structures, where O-Ac is O-acetylation of Neu5Ac. Neu5AcNeu5Ac is most likely Neu5Ac 2,8Neu5Ac linked. O-glycosylations in the vicinity of the cleavage sites may influence the proteolytic processing. Appicans are L-APP isoforms with O-linked chondroitin sulfate. Phosphorylation in the C-terminal on tyrosine, threonine and serine residues is neuron-specific. Phosphorylation can affect APP processing, neuronal differentiation and interaction with other proteins. Phosphorylated on Thr-743 in neuronal cells by Cdc5 kinase and Mapk10, in dividing cells by Cdc2 kinase in a cell-cycle dependent manner with maximal levels at the G2/M phase and, in vitro, by GSK-3-beta. The Thr-743 phosphorylated form causes a conformational change which reduces binding of Fe65 family members. In dopaminergic (DA) neurons, phosphorylation on Thr-743 by LRKK2 promotes the production and the nuclear translocation of the APP intracellular domain (AICD) which induces DA neuron apoptosis. Phosphorylation on Tyr-757 is required for SHC binding. Phosphorylated in the extracellular domain by casein kinases on both soluble and membrane-bound APP. This phosphorylation is inhibited by heparin. Extracellular binding and reduction of copper, results in a corresponding oxidation of Cys-144 and Cys-158, and the formation of a disulfide bond. In vitro, the APP-Cu(+) complex in the presence of hydrogen peroxide results in an increased production of amyloid-beta-containing peptides. Trophic-factor deprivation triggers the cleavage of surface APP by beta-secretase to release sAPP-beta which is further cleaved to release an N-terminal fragment of APP (N-APP). Amyloid-beta peptides are degraded by IDE. Sulfated on tyrosine residues. |
| Function | Functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Interaction between APP molecules on neighboring cells promotes synaptogenesis. Involved in cell mobility and transcription regulation through protein-protein interactions. Can promote transcription activation through binding to APBB1-KAT5 and inhibits Notch signaling through interaction with Numb. Couples to apoptosis-inducing pathways such as those mediated by G(o) and JIP. Inhibits G(o) alpha ATPase activity. Acts as a kinesin I membrane receptor, mediating the axonal transport of beta-secretase and presenilin 1. By acting as a kinesin I membrane receptor, plays a role in axonal anterograde transport of cargo towards synapses in axons. Involved in copper homeostasis/oxidative stress through copper ion reduction. In vitro, copper-metallated APP induces neuronal death directly or is potentiated through Cu(2+)-mediated low-density lipoprotein oxidation. Can regulate neurite outgrowth through binding to components of the extracellular matrix such as heparin and collagen I and IV. The splice isoforms that contain the BPTI domain possess protease inhibitor activity. Induces a AGER-dependent pathway that involves activation of p38 MAPK, resulting in internalization of amyloid-beta peptide and leading to mitochondrial dysfunction in cultured cortical neurons. Provides Cu(2+) ions for GPC1 which are required for release of nitric oxide (NO) and subsequent degradation of the heparan sulfate chains on GPC1. Amyloid-beta peptides are lipophilic metal chelators with metal-reducing activity. Bind transient metals such as copper, zinc and iron. In vitro, can reduce Cu(2+) and Fe(3+) to Cu(+) and Fe(2+), respectively. Amyloid-beta protein 42 is a more effective reductant than amyloid-beta protein 40. Amyloid-beta peptides bind to lipoproteins and apolipoproteins E and J in the CSF and to HDL particles in plasma, inhibiting metal-catalyzed oxidation of lipoproteins. APP42-beta may activate mononuclear phagocytes in the brain and elicit inflammatory responses. Promotes both tau aggregation and TPK II-mediated phosphorylation. Interaction with overexpressed HADH2 leads to oxidative stress and neurotoxicity. Also binds GPC1 in lipid rafts. Appicans elicit adhesion of neural cells to the extracellular matrix and may regulate neurite outgrowth in the brain. The gamma-CTF peptides as well as the caspase-cleaved peptides, including C31, are potent enhancers of neuronal apoptosis. N-APP binds TNFRSF21 triggering caspase activation and degeneration of both neuronal cell bodies (via caspase-3) and axons (via caspase-6). |
| Protein Name | Amyloid-Beta Precursor ProteinAppAbppAppiAlzheimer Disease Amyloid A4 Protein HomologAlzheimer Disease Amyloid ProteinAmyloid Precursor ProteinAmyloid-BetaA4 Precursor ProteinAmyloid-Beta A4 ProteinCerebral Vascular Amyloid PeptideCvapPrea4Protease Nexin-IiPn-Ii Cleaved Into - N-App - Soluble App-AlphaS-App-Alpha - Soluble App-BetaS-App-Beta - C99Beta-Secretase C-Terminal FragmentBeta-Ctf - Amyloid-Beta Protein 42Abeta42Beta-App42 - Amyloid-Beta Protein 40Abeta40Beta-App40 - C83Alpha-Secretase C-Terminal FragmentAlpha-Ctf - P3(42 - P3(40 - C80 - Gamma-Secretase C-Terminal Fragment 59Amyloid Intracellular Domain 59Aicd-59Aid(59Gamma-Ctf(59 - Gamma-Secretase C-Terminal Fragment 57Amyloid Intracellular Domain 57Aicd-57Aid(57Gamma-Ctf(57 - Gamma-Secretase C-Terminal Fragment 50Amyloid Intracellular Domain 50Aicd-50Aid(50Gamma-Ctf(50 - C31 |
| Database Links | Reactome: R-HSA-114608Reactome: R-HSA-3000178Reactome: R-HSA-381426Reactome: R-HSA-416476Reactome: R-HSA-418594Reactome: R-HSA-432720Reactome: R-HSA-444473Reactome: R-HSA-445989Reactome: R-HSA-844456Reactome: R-HSA-879415Reactome: R-HSA-8862803Reactome: R-HSA-8957275Reactome: R-HSA-933542Reactome: R-HSA-9609523Reactome: R-HSA-9660826Reactome: R-HSA-977225 |
| Cellular Localisation | Cell MembraneSingle-Pass Type I Membrane ProteinMembranePerikaryonCell ProjectionGrowth ConeClathrin-Coated PitEarly EndosomeCytoplasmic VesicleCell Surface Protein That Rapidly Becomes Internalized Via Clathrin-Coated PitsOnly A Minor Proportion Is Present At The Cell MembraneMost Of The Protein Is Present In Intracellular VesiclesDuring MaturationThe Immature App (N-Glycosylated In The Endoplasmic Reticulum) Moves To The Golgi Complex Where Complete Maturation Occurs (O-Glycosylated And Sulfated)After Alpha-Secretase CleavageSoluble App Is Released Into The Extracellular Space And The C-Terminal Is Internalized To Endosomes And LysosomesSome App Accumulates In Secretory Transport Vesicles Leaving The Late Golgi Compartment And Returns To The Cell SurfaceApp Sorts To The Basolateral Surface In Epithelial CellsDuring Neuronal DifferentiationThe Thr-743 Phosphorylated Form Is Located Mainly In Growth ConesModerately In Neurites And Sparingly In The Cell BodyCasein Kinase Phosphorylation Can Occur Either At The Cell Surface Or Within A Post-Golgi CompartmentAssociates With Gpc1 In Perinuclear CompartmentsColocalizes With Sorl1 In A Vesicular Pattern In Cytoplasm And Perinuclear RegionsC83: Endoplasmic ReticulumGolgi ApparatusC99: Early EndosomeSoluble App-Beta: SecretedAmyloid-Beta Protein 40: Cell SurfaceAmyloid-Beta Protein 42: Cell SurfaceAssociates With Fpr2 At The Cell Surface And The Complex Is Then Rapidly InternalizedGamma-Secretase C-Terminal Fragment 59: NucleusCytoplasmLocated To Both The Cytoplasm And Nuclei Of NeuronsIt Can Be Translocated To The Nucleus Through Association With Apbb1 (Fe65)In Dopaminergic NeuronsThe Phosphorylated Thr-743 Form Is Localized To The Nucleus |
| Alternative ELISA Names | Amyloid-Beta Precursor Protein ELISA kitApp ELISA kitAbpp ELISA kitAppi ELISA kitAlzheimer Disease Amyloid A4 Protein Homolog ELISA kitAlzheimer Disease Amyloid Protein ELISA kitAmyloid Precursor Protein ELISA kitAmyloid-Beta ELISA kitA4 Precursor Protein ELISA kitAmyloid-Beta A4 Protein ELISA kitCerebral Vascular Amyloid Peptide ELISA kitCvap ELISA kitPrea4 ELISA kitProtease Nexin-Ii ELISA kitPn-Ii Cleaved Into - N-App - Soluble App-Alpha ELISA kitS-App-Alpha - Soluble App-Beta ELISA kitS-App-Beta - C99 ELISA kitBeta-Secretase C-Terminal Fragment ELISA kitBeta-Ctf - Amyloid-Beta Protein 42 ELISA kitAbeta42 ELISA kitBeta-App42 - Amyloid-Beta Protein 40 ELISA kitAbeta40 ELISA kitBeta-App40 - C83 ELISA kitAlpha-Secretase C-Terminal Fragment ELISA kitAlpha-Ctf - P3(42 - P3(40 - C80 - Gamma-Secretase C-Terminal Fragment 59 ELISA kitAmyloid Intracellular Domain 59 ELISA kitAicd-59 ELISA kitAid(59 ELISA kitGamma-Ctf(59 - Gamma-Secretase C-Terminal Fragment 57 ELISA kitAmyloid Intracellular Domain 57 ELISA kitAicd-57 ELISA kitAid(57 ELISA kitGamma-Ctf(57 - Gamma-Secretase C-Terminal Fragment 50 ELISA kitAmyloid Intracellular Domain 50 ELISA kitAicd-50 ELISA kitAid(50 ELISA kitGamma-Ctf(50 - C31 ELISA kitAPP ELISA kitA4 ELISA kitAD1 ELISA kit |
| Specificity | This kit recognizes Human Aβ1-40 in samples. No significant cross-reactivity or interference between Human Aβ1-40 and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
|---|---|---|
| Micro ELISA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent | 1 vial, 10 mL | 2-8℃ (Protect from light) |
| Stop Solution | 1 vial, 10 mL | 2-8°C |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy |
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 93-109 | 101 |
| EDTA plasma (n=8) | 85-96 | 91 |
| Cell culture media (n=8) | 92-108 | 100 |
| Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision | |
|---|---|---|---|---|---|---|
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (pg/mL) | 50.60 | 152.10 | 454.70 | 45.80 | 156.80 | 436.80 |
| Standard deviation | 3.30 | 6.10 | 23.60 | 2.60 | 9.10 | 13.10 |
| CV (%) | 6.52 | 4.01 | 5.19 | 5.68 | 5.80 | 3.00 |
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance
