Anti-Amyloid Precursor Protein (phospho Y757) antibody (ab19850)

Overview

  • Product nameAnti-Amyloid Precursor Protein (phospho Y757) antibodySee all Amyloid Precursor Protein primary antibodies ...
  • Description
    Rabbit polyclonal to Amyloid Precursor Protein (phospho Y757)
  • SpecificityRabbit polyclonal to APP (phospho Y757) ab19850 recognises various isoforms of APP in the phosphorylated state. ab19850 recognises both the mature (around 80 kDa) and immature (around 110 kDa) isoforms of APP. 10 named APP isoforms exist following alternative splicing, as well as a number of additional isoforms for which experimental confirmation may be lacking. Several C-terminal APP fragments around 50 kDa also exist and are well documented (Swiss Prot, Haverster EMBL databases).
  • Tested applicationsWB, IHC-FoFr more details
  • Species reactivity
    Reacts with: Mouse
    Predicted to work with: Rat, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 750 to the C-terminus of Human Amyloid Precursor Protein, phosphorylated at Y757.

    (Peptide available as ab20648.)

  • Positive control
    • mouse brain
  • General notes


    Phosphorylation of APP at Tyr757 and 762 is important for MAPK8IP1, APBA1, shcA/shcC and DAB1 binding. Binding interactions regulated by phosphorylation determine the localisation and the function of APP. Therefore Amyloid Precursor Protein (phospho Y757) antibody will be a useful tool in tracing the processing, distribution and interaction of APP.

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Primary antibody notes Phosphorylation of APP at Tyr757 and 762 is important for MAPK8IP1, APBA1, shcA/shcC and DAB1 binding. Binding interactions regulated by phosphorylation determine the localisation and the function of APP. Therefore Amyloid Precursor Protein (phospho Y757) antibody will be a useful tool in tracing the processing, distribution and interaction of APP.
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab19850 in the following tested applications.

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

Application Notes
WB
IHC-FoFr
  • Application notesIHC-Fo: 1/1000.
    WB: Use at a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 55, 87, 110, 170 kDa (predicted molecular weight: 55, 87, 110 kDa).

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionFunctions 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 Also bind 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 specificityExpressed 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 diseaseAlzheimer disease 1
      Cerebral amyloid angiopathy, APP-related
    • Sequence similaritiesBelongs to the APP family.
      Contains 1 BPTI/Kunitz inhibitor domain.
    • DomainThe 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 terminatiing 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 localizationMembrane. 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
    • Database links
    • Alternative names
      • A4 amyloid protein antibody
      • A4 antibody
      • A4_HUMAN antibody
      • AAA antibody
      • ABETA antibody
      • ABPP antibody
      • AD 1 antibody
      • AD1 antibody
      • AICD-50 antibody
      • AICD-57 antibody
      • AICD-59 antibody
      • AID(50) antibody
      • AID(57) antibody
      • AID(59) antibody
      • Alzheimer disease 1 antibody
      • Alzheimer disease amyloid protein antibody
      • Alzheimer disease antibody
      • Alzheimer's disease amyloid protein antibody
      • Amyloid beta (A4) precursor protein antibody
      • Amyloid beta A4 protein antibody
      • Amyloid beta A4 protein precursor isoform b antibody
      • Amyloid beta A4 protein precursor isoform b antibody
      • Amyloid beta A4 protein precursor isoform c antibody
      • Amyloid beta A4 protein precursor isoform c antibody
      • Amyloid beta A4 protein precursor isoform a antibody
      • Amyloid beta A4 protein precursor isoform b antibody
      • Amyloid beta A4 protein precursor isoform b antibody
      • Amyloid beta A4 protein precursor isoform c antibody
      • Amyloid beta A4 protein precursor isoform c antibody
      • Amyloid beta protein antibody
      • Amyloid beta-peptide antibody
      • Amyloid intracellular domain 50 antibody
      • Amyloid intracellular domain 57 antibody
      • Amyloid intracellular domain 59 antibody
      • Amyloid of aging and alzheimer disease antibody
      • APP antibody
      • APP I antibody
      • APPI antibody
      • Beta amyloid peptide antibody
      • Beta-APP40 antibody
      • Beta-APP42 antibody
      • C31 antibody
      • Cerebral vascular amyloid peptide antibody
      • CTFgamma antibody
      • CVAP antibody
      • Gamma-CTF(50) antibody
      • Gamma-CTF(57) antibody
      • Gamma-CTF(59) antibody
      • Human mRNA for amyloid A4 precursor of Alzheimer's disease antibody
      • peptidase nexin II antibody
      • PN 2 antibody
      • PN II antibody
      • PN-II antibody
      • PN2 antibody
      • PreA4 antibody
      • Protease nexin II antibody
      • Protease nexin-II antibody
      • S-APP-alpha antibody
      • S-APP-beta antibody
      see all

    Anti-Amyloid Precursor Protein (phospho Y757) antibody images

    • Lane 1 : Amyloid Precursor Protein (non modified) C-terminal antibody
      Lane 2 : Anti-Amyloid Precursor Protein (phospho Y757) antibody (ab19850) at 0.5 µg/ml

      Lane 1 : mouse dentate gyrus lysate
      Lane 2 : mouse dentate gyrus lysate

      Secondary
      Rabbit IgG 2nd ab at 1/25000 dilution

      Predicted band size : 55, 87, 110 kDa
      Observed band size : 55,87,110 kDa
      Additional bands at : 170 kDa (possible isoform).

      Florian Plattner, Wolfson Inst Biomed Research, United Kingdom

    • Anti-Amyloid Precursor Protein (phospho Y757) antibody (ab19850) at 1/1000 dilution + Mouse Hippocampus whole cell lysate

      Secondary
      Goat anti-rabbit IgG HRP
      developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 55, 87, 110 kDa
      Observed band size : 50,95,130 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 170 kDa,190 kDa (possible non-specific binding). We are unsure as to the identity of these extra bands.

      Exposure time : 1 minute

      See Abreview

    • ab19850 at 1/1000 staining mouse brain tissue sections by IHC (Formalin/PFA fixed sections). The tissue was paraformaldehyde fixed and a heat mediated antigen retrieval step in a Na Citrate buffer was performed. The tissue was permeabilized and incubated with ab19850 for 18 hours. A biotinylated goat anti-rabbit IgG was used as the secondary. The sections were counterstained with Congo red. The image shows Congo red positive plaque core surrounded by phospho-APP positive neurites in AD model mouse.

      See Abreview

    References for Anti-Amyloid Precursor Protein (phospho Y757) antibody (ab19850)

    ab19850 has not yet been referenced specifically in any publications.

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    The possible explanation for thedifference in molecular weightcould bedue to the difference in glycosylaiton of the protein. You may check publications to know more about this difference.

    I hope this i...

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    The sequence is present in all isoforms except the second one:Isoform APP305 as 306-770aa is missing. Altogether 10 isoforms (http://www.uniprot.org/uniprot/P05067)

    Application Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample Mouse Tissue sections (brain)
    Specification brain
    Fixative Paraformaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Na Citrate sol. pH=6.0 for 30 min
    Permeabilization Yes - Triton 100X
    Username

    Dr. Inga Kadisha

    Verified customer

    Submitted Jun 01 2007

    Application Western blot
    Sample Mouse Tissue lysate - whole (Hippocampus)
    Specification Hippocampus
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
    Username

    Mr. Florian Plattner

    Verified customer

    Submitted Dec 07 2005

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"