Overview

  • Product name
    Anti-Amyloid Precursor Protein antibody
    See all Amyloid Precursor Protein primary antibodies
  • Description
    Rabbit polyclonal to Amyloid Precursor Protein
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Monkey
  • Immunogen

    Synthetic peptide corresponding to Human Amyloid Precursor Protein aa 737-731 (C terminal).
    Sequence:

    GYENPTYKFFEQMQN


    (Peptide available as ab7874)

  • Positive control
    • Murine brain tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab2073 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
ICC/IF Use a concentration of 20 µg/ml.
WB 1/500 - 1/1000. Can be blocked with Amyloid Precursor Protein peptide (ab7874).
IHC-P Use at an assay dependent concentration.

Target

  • 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • A4 amyloid protein antibody
    • A4_HUMAN antibody
    • AAA antibody
    • ABETA antibody
    • ABPP antibody
    • AD1 antibody
    • AICD-50 antibody
    • AICD-57 antibody
    • AICD-59 antibody
    • AID(50) antibody
    • AID(57) antibody
    • AID(59) antibody
    • Alzheimer disease amyloid protein antibody
    • Amyloid beta (A4) precursor protein antibody
    • Amyloid beta A4 protein antibody
    • Amyloid beta protein antibody
    • Amyloid intracellular domain 50 antibody
    • Amyloid intracellular domain 57 antibody
    • Amyloid intracellular domain 59 antibody
    • Amyloid precursor protein antibody
    • APP antibody
    • APPI antibody
    • Beta amyloid peptide antibody
    • Beta-amyloid precursor protein 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
    • 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

Images

  • Immunohistochemistry (Formalin-fixed paraffin embedded sections) of mouse brain tissue labeling Amyloid Precursor Protein with Anti-Amyloid Precursor Protein antibody (ab2073) at 5μg/ml.
  • Western Blot of mouse brain lysate labeling Amyloid Precursor Protein with Anti-Amyloid Precursor Protein antibody (ab2073) at (1) 0.5 and (2) 1µg/ml.
  • Western blot analysis of APP in mouse (M) and rat (R) brain tissue lysates with anti-APP (CT) at 1:500 dilution. Western blot analysis of APP in mouse (M) and rat (R) brain tissue lysates with anti-APP (CT) at 1:500 dilution.
  • Immunohistochemical staining (paraffin embedded) of rat brain using anti-APP at 2 µg/ml.
  • Immunofluorescence of APP in Rat Brain cells using ab2073 at 20 ug/ml.

References

This product has been referenced in:
  • Brai E  et al. Notch1 hallmarks fibrillary depositions in sporadic Alzheimer's disease. Acta Neuropathol Commun 4:64 (2016). Read more (PubMed: 27364742) »
  • Gavín R  et al. Involvement of Dab1 in APP processing and beta-amyloid deposition in sporadic Creutzfeldt-Jakob patients. Neurobiol Dis 37:324-9 (2010). IHC-P, IHC-Fr ; Human . Read more (PubMed: 19853035) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for your reply. We are confident that our product works properly. However, we are willing to offer you a complementary vial of our other anti-APP antibody, catalog number ab2072. This one is against an N-terminal peptide of APP. Please do let us know if you would like to try it or not. If yes, would you be so kind to confirm your Purchase Order Number or Abcam Order Number please.

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Answer

Thank you for your response. If you don't see the desired band: 1. It may be due to inadequate transfer of proteins during electroblotting. Check our western blot protocol to optimize your electrophoresis and transfer conditions. 2. You could increase the concentration of the primary antibody, such as 1:500 or 1:250, as well as the secondary antibody. Also, confirm that you have a highly sensitive secondary antibody. 3. You could try longer exposure of the membrane to X-ray film (30 minutes to over night). 4. The detection enzyme may be inactivated. Sodium azide inactivates horseradish peroxidase (HRP) irreversibly. Also, bacterial contamination also diminishes HRP activity. 5. Polyvinyl wrap from some sources has been found to hide the signal. You could repeat the incubation with chemiluminescence reagents and place the blot between two pieces of acetate transparency film. 6. You should try using a lysate that is known to express the protein of interest as a positive control. It is important to know if the cells or tissue express the protein of interest. Most proteins are not expressed in all kinds of cells or tissues. We can offer you a positive control for the anti-APP antibody. This is the mouse brain tissue lysate, catalog number ab7934. Please do let us know if you would like to get one vial.

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Question

I order a vial of (1 x ab2073) Rabbit polyclonal to Amyloid Precursor Protein and received the product on Jun 17th, 2004. I divided the product (lot 55621) into aliquots sand stored them at -20 degree as suggested. I used this antibody to perform the western blot but never worked. I wanted to know what happened to the ab2073. How did I do the western blot? Here it goes. (1) Protein was overexpressed in both CHO and Hek193 cells (2) After transferred from gel to NC membr, NC membr was incubated with 1:1000 diluted ab2073, room temperature, 1.0 hr As control, two separated NC memebrs were incubated with anti-his and anti-myc antibodies, room temperature, 1.0 hr. (3) After washed several times, NC membr was incubated with 1:50 000 diluted monoclonal anti-rabbit immunoglulins AP conjugate clone RG-16 (Sigma, product No A2306, lot 073K4827), room temperature, 1.0 hr. Meanwhile, the other two membrs were incubated with goat anti-mouse secondary antibody AP conjugate, room temperature, 1.0 hr (4) After stained by substrate NCIP/NBT, no bands showed up on anti-APP antibody incubated membr. Meanwhile, the other two gave strong bands. In order to improve the anti-APP antibody system, I tried several times, never suceeded. I also tried to prolong the incubation time and raise the dilution ratio. Here is some details. Raising dilution ratio: (1) Incubate the transferred NC membr with 1:500 ab2073, room temperature, 1.0 hr; (2) After washed several times, membr was incubated with 1:20 000 A2306, room temperature, 1.5 hr; (3) After stained with substrate, NO bands showed up on NC membr. Prolonging incubation time (1) Incubate the transferred NC membr with 1:500 ab2073, for 21 hrs at 4 degree; (2) After washed, NC membr was incubated with 1:1000 A2306, for 2 hr room temperature. (3) after staining with substrate, NO bands showed up. Meanwhile, anti-his (or anti-myc) antibody systems gave strong bands on the same condition. I don't know why this happened. Is it possible that ab2073 is not in good condition. Or the quality is not so good? If so, may we get the refund or ... I am really in hurry, beacause we have a deadline to meet. Though I spent a lot of time and effort on this ab2073 antibody system, I didn't achieve any success.

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Answer

Thank you for your enquiry. At what terminus are the tags? The antibody was raised against a peptide sequence at the C-terminus and it is possible that the tags are inhibiting the binding of the antibody. This is probably why the anti-his and anti-myc antibodies provide results while the anti-APP does not. The antibody recognizes the APP precursor and not the cleaved form of APP. The peptide sequence used for immunization is from C99, which is cleaved from the APP precursor by beta-secretase. This antibody has been tested for Western blot using murine brain tissue lysate. We would suggest running a positive control along with the samples.

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Question
Answer

The antibody to our knowledge has not yet been tested in IP. Immediately we have done this or heard that it does work in this application we will publish it on the datasheet.

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Question
Answer

Unfortunatetly, this antibody has only been tested by western blot. It probably will work in FFPE sections but we can't say for sure until it has been tested

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

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