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Neuroscience Neurology process Neurodegenerative disease Alzheimer's disease Amyloid
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Recombinant

Human Amyloid Precursor Protein Matched Antibody Pair Kit (ab220128)

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  • Protocol Booklet
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Human Amyloid Precursor Protein standard curve
  • Sandwich ELISA - Human Amyloid Precursor Protein Matched Antibody Pair Kit (ab220128)

Key features and details

  • Unlabeled capture antibody, biotin-labeled detection antibody and calibrated protein standard
  • For economical ELISA and ELISA-based assay development
  • Reacts with: Human
  • Range: 125 pg/ml - 8000 pg/ml

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Overview

  • Product name

    Human Amyloid Precursor Protein Matched Antibody Pair Kit
    See all Amyloid Precursor Protein kits
  • Detection method

    Colorimetric
  • Assay type

    ELISA set
  • Sensitivity

    32.78 pg/ml
  • Range

    125 pg/ml - 8000 pg/ml
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human Amyloid Precursor Protein Matched Antibody Pair Kits include a capture and a biotinylated detector antibody pair, along with a calibrated protein standard, suitable for sandwich ELISA. The Matched Antibody Pair Kit can be used to quantify native and recombinant human Amyloid Precursor Protein.


    Optimization of the kit reagents to sample type, immunoassay format or instrumentation may be required. Guidelines for use of this kit in a standard 96-well microplate sandwich ELISA using HRP/TMB system of colorimetric detection is described in this assay procedure for the purposes of quantification.


    Protocol information and tips on the use of the Matched Antibody Pair kits for sandwich ELISA can be found on our website. An accessory pack can be purchased which includes buffer reagents required to perform 10 x 96-well plate sandwich ELISAs (ab210905).


    For additional information on the performance of the antibody pair used in this kit, please see our equivalent SimpleStep ELISA kit ab216944. Please note that while the antibody pair is the same provided in the corresponding SimpleStep ELISA Kit, due to differences in their formulation, this antibody pair cannot be used with the consumables provided with our SimpleStep ELISA Kits.

  • Tested applications

    Suitable for: ELISAmore details
  • Platform

    Reagents

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 10 x 96 tests 5 x 96 tests
    Human Amyloid Precursor Protein Capture Antibody 2 x 50µg 1 x 50µg
    Human Amyloid Precursor Protein Detector Antibody 2 x 12.5µg 1 x 12.5µg
    Human Amyloid Precursor Protein Lyophilized Protein 2 vials 1 vial
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Amyloid
  • Function

    Functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. 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 (By similarity). Acts as a kinesin I membrane receptor, mediating the axonal transport of beta-secretase and presenilin 1. 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.
    Beta-amyloid 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. Beta-amyloid 42 is a more effective reductant than beta-amyloid 40. Beta-amyloid peptides bind to lipoproteins and apolipoproteins E and J in the CSF and to HDL particles in plasma, inhibiting metal-catalyzed oxidation of lipoproteins. Beta-APP42 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).
  • Tissue specificity

    Expressed in all fetal tissues examined with highest levels in brain, kidney, heart and spleen. Weak expression in liver. In adult brain, highest expression found in the frontal lobe of the cortex and in the anterior perisylvian cortex-opercular gyri. Moderate expression in the cerebellar cortex, the posterior perisylvian cortex-opercular gyri and the temporal associated cortex. Weak expression found in the striate, extra-striate and motor cortices. Expressed in cerebrospinal fluid, and plasma. Isoform APP695 is the predominant form in neuronal tissue, isoform APP751 and isoform APP770 are widely expressed in non-neuronal cells. Isoform APP751 is the most abundant form in T-lymphocytes. Appican is expressed in astrocytes.
  • Involvement in disease

    Alzheimer disease 1
    Cerebral amyloid angiopathy, APP-related
  • Sequence similarities

    Belongs to the APP family.
    Contains 1 BPTI/Kunitz inhibitor domain.
  • Domain

    The basolateral sorting signal (BaSS) is required for sorting of membrane proteins to the basolateral surface of epithelial cells.
    The NPXY sequence motif found in many tyrosine-phosphorylated proteins is required for the specific binding of the PID domain. However, additional amino acids either N- or C-terminal to the NPXY motif are often required for complete interaction. The PID domain-containing proteins which bind APP require the YENPTY motif for full interaction. These interactions are independent of phosphorylation on the terminal tyrosine residue. The NPXY site is also involved in clathrin-mediated endocytosis.
  • 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 40 (Abeta40) and amyloid-beta 42 (Abeta42), major components of amyloid plaques, and the cytotoxic C-terminal fragments, gamma-CTF(50), gamma-CTF(57) and gamma-CTF(59). Many other minor beta-amyloid peptides, beta-amyloid 1-X peptides, are found in cerebral spinal fluid (CSF) including the beta-amyloid 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 caspase-6, -8 or -9 results in the production of the neurotoxic C31 peptide and the increased production of beta-amyloid peptides.
    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 beta-amyloid peptides (beta-amyloid 1-15, 1-16, 1-17, 1-18, 1-19 and 1-20) but not found on beta-amyloid 38, beta-amyloid 40 nor on beta-amyloid 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. 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 beta-amyloid-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).
    Beta-amyloid peptides are degraded by IDE.
  • Cellular localization

    Membrane. Membrane, clathrin-coated pit. Cell surface protein that rapidly becomes internalized via clathrin-coated pits. During maturation, the immature APP (N-glycosylated in the endoplasmic reticulum) moves to the Golgi complex where complete maturation occurs (O-glycosylated and sulfated). After alpha-secretase cleavage, soluble APP is released into the extracellular space and the C-terminal is internalized to endosomes and lysosomes. Some APP accumulates in secretory transport vesicles leaving the late Golgi compartment and returns to the cell surface. Gamma-CTF(59) peptide is located to both the cytoplasm and nuclei of neurons. It can be translocated to the nucleus through association with APBB1 (Fe65). Beta-APP42 associates with FRPL1 at the cell surface and the complex is then rapidly internalized. APP sorts to the basolateral surface in epithelial cells. During neuronal differentiation, the Thr-743 phosphorylated form is located mainly in growth cones, moderately in neurites and sparingly in the cell body. Casein kinase phosphorylation can occur either at the cell surface or within a post-Golgi compartment. Associates with GPC1 in perinuclear compartments. Colocalizes with SORL1 in a vesicular pattern in cytoplasm and perinuclear regions.
  • Target information above from: UniProt accession P05067 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • A4 amyloid protein
    • A4_HUMAN
    • AAA
    • ABETA
    • ABPP
    • AD1
    • AICD-50
    • AICD-57
    • AICD-59
    • AID(50)
    • AID(57)
    • AID(59)
    • Alzheimer disease amyloid protein
    • Amyloid beta (A4) precursor protein
    • Amyloid beta A4 protein
    • Amyloid beta protein
    • Amyloid intracellular domain 50
    • Amyloid intracellular domain 57
    • Amyloid intracellular domain 59
    • Amyloid precursor protein
    • APP
    • APPI
    • Beta amyloid peptide
    • beta-amyloid peptide
    • Beta-amyloid precursor protein
    • Beta-APP40
    • Beta-APP42
    • C31
    • Cerebral vascular amyloid peptide
    • CTFgamma
    • CVAP
    • Gamma-CTF(50)
    • Gamma-CTF(57)
    • Gamma-CTF(59)
    • peptidase nexin II
    • peptidase nexin-II
    • PN 2
    • PN II
    • PN-II
    • PN2
    • PreA4
    • Protease nexin II
    • Protease nexin-II
    • S-APP-alpha
    • S-APP-beta
    • sAPP
    see all
  • Database links

    • Entrez Gene: 351 Human
    • Omim: 104760 Human
    • SwissProt: P05067 Human
    • Unigene: 434980 Human

    Associated products

    • Alternative Versions

      • Human Amyloid Precursor Protein Antibody Pair - BSA and Azide free (ab241866)
    • Related Products

      • ELISA Accessory Pack (ab210905)
    • SimpleStep ELISA kits

      • Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Applications

    Our Abpromise guarantee covers the use of ab220128 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    ELISA Use at an assay dependent concentration.

    Images

    • Human Amyloid Precursor Protein standard curve
      Human Amyloid Precursor Protein standard curve

      Standard calibration curve. Background subtracted values are graphed.

    • Sandwich ELISA - Human Amyloid Precursor Protein Matched Antibody Pair Kit (ab220128)
      Sandwich ELISA - Human Amyloid Precursor Protein Matched Antibody Pair Kit (ab220128)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab220128? Please let us know so that we can cite the reference in this datasheet.

    ab220128 has not yet been referenced specifically in any publications.

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