RabMAb

Anti-Amyloid beta precursor protein antibody [Y188] (ab32136)

Overview

  • Product nameAnti-Amyloid beta precursor protein antibody [Y188]See all Amyloid beta precursor protein primary antibodies ...
  • Description
    Rabbit monoclonal [Y188] to Amyloid beta precursor protein
  • Tested applicationsWB, IHC-P, ICC, Flow Cyt, IP, IHC-FoFr, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to A synthetic peptide corresponding to the C-terminus of human APP.

  • Positive control
    • WB: Hela cell lysate. IHC-P: Human brain tissue.
  • General notesProduced under U.S. Patent No. 5,675,063.

Properties

Applications

Our Abpromise guarantee covers the use of ab32136 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
WB 1/100 - 1/10000. Detects a band of approximately 95 kDa (predicted molecular weight: 87 kDa).
IHC-P Use at an assay dependent dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC 1/250 - 1/500.
Flow Cyt 1/100.
IP 1/50.
IHC-FoFr 1/750. PubMed: 18974297
IHC-Fr Use at an assay dependent concentration.

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.
  • Information by UniProt
  • Database links
  • FormThere are 10 isoforms produced by alternative splicing. Amyloid beta precursor protein is cleaved into the following 14 chains: 1)N-APP2, 2)Soluble APP-alpha, 3)Soluble APP-beta, 4)C99, 5)Beta-amyloid protein 42, 6) Beta-amyloid protein 40, 70 C83, 80 P3,(42) 90 P3(40), 10) C80, 11) Gamma-secretase C-terminal fragment 59, 12) Gamma-secretase C-terminal fragment 57, 13) Gamma-secretase C-terminal fragment 50, 14) C31.
  • 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's Disease Amyloid Protein antibody
    • Amyloid beta (A4) precursor protein antibody
    • Amyloid beta A4 protein antibody
    • Amyloid Beta A4 Protein Precursor Isoform A antibody
    • Amyloid Beta A4 Protein Precursor Isoform B antibody
    • Amyloid Beta A4 Protein Precursor Isoform C antibody
    • Amyloid Beta Peptide antibody
    • Amyloid beta protein 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
    • APPI antibody
    • Beta amyloid peptide antibody
    • Beta-APP40 antibody
    • Beta-APP42 antibody
    • C31 antibody
    • Cerebal Vascular Amyloid Peptide antibody
    • Cerebral vascular amyloid peptide antibody
    • CTFgamma 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
    • peptidase nexin-II antibody
    • PN 2 antibody
    • PN II antibody
    • PN-II antibody
    • PN2 antibody
    • PNII antibody
    • PREA4 antibody
    • Protease Nexin II antibody
    • Protease nexin-II antibody
    • S-APP-alpha antibody
    • S-APP-beta antibody
    see all

Anti-Amyloid beta precursor protein antibody [Y188] images

  • Anti-Amyloid beta precursor protein antibody [Y188] (ab32136) at 1/20000 dilution + Hela cell lysate

    Predicted band size : 87 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 110 kDa. We are unsure as to the identity of these extra bands.
  • Ab32136, at a 1/250 dilution, staining Amyloid beta precursor protein by immunohistochemistry.
    Positive immunohistochemical staining, using paraffin embedded human brain tissue (A).
    Negative immunohistochemical staining, using human breast (B), skeletal muscle (C) and liver (D) tissues.
    Tissues were stained in parallel on the same Normal Tissue Array.
  • ab32136, staining Amyloid beta precursor protein (green) in spinal cords from wild-type (left) or APP-/- (right) mouse embryos by Immunohistochemistry (PFA perfusion fixed frozen sections).

    Embryos were fixed overnight in 4% paraformaldehyde at 4°C, permeabilized for 30 min in PBS containing 0.1% Triton X-100 and blocked for 4 h in PBS containing 3% normal donkey serum, 2% BSA, and 0.1% Triton X-100. Sections were incubated overnight at 4 °C with anti-APP antibody at 1/200 dilution. An AlexaFluor®488-conjugated donkey anti-rabbit IgG (1/200) was used as the secondary antibody.
  • Anti-Amyloid beta precursor protein antibody [Y188] (ab32136) at 1/1000 dilution + Mouse spleen whole cell lysate at 100 µg

    Secondary
    HRP-conjugated goat anti-rabbit polyclonal IgG at 1/4000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 87 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 125 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Overlay histogram showing PC12 cells stained with ab32136 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32136, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in PC12 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-Amyloid beta precursor protein antibody [Y188] (ab32136)

This product has been referenced in:
  • Villegas C  et al. Dual-tagged amyloid-ß precursor protein reveals distinct transport pathways of its N- and C-terminal fragments. Hum Mol Genet 23:1631-43 (2014). Read more (PubMed: 24203698) »
  • Olsen O  et al. Genetic analysis reveals that amyloid precursor protein and death receptor 6 function in the same pathway to control axonal pruning independent of ß-secretase. J Neurosci 34:6438-47 (2014). Read more (PubMed: 24806670) »

See all 15 Publications for this product

Product Wall

The following email which contains your credit note id and codes for two free primary antibodies was sent to you last night. I am sorry if this did not arrive and hope that any technical issue has been resolved. Please contact me should you have furthe...

Read More

Thank you for contacting us.

We do offer a number of anti-APP antibodies which are validated and guaranteed to work in western blot in mouse and human. Three of these which have the most data available are ab12269, ab32136, ab78314. These ...

Read More

Thank you for contacting us.

Your credit note ID is *.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask y...

Read More

Thank you for your reply. Each credit note has a value of xxxx. We ask that you please contact our Finance department at mailto:creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corne...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse spleen whole extract)
Loading amount 100 µg
Specification Mouse spleen whole extract
Gel Running Conditions Reduced Denaturing (Tris-Glycine -10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 01 2012

Thank you for your reply, I am glad you have found the information helpful. I am only sorry for the initial confusion caused.
I have now arranged for the information regarding the specificity of the antibody to be sent out to previous customers of...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse spleen whole extract)
Loading amount 100 µg
Specification Mouse spleen whole extract
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 30 2012

Thank you for your reply, and sorry for the delay in getting back to you.
I now have some further information to share with you in regards to the Anti-Amyloid beta precursor protein antibody (ab32136).
I have had confirmation that the antibod...

Read More

Thank you very much for that information.
I can now understand why we could not see the same information on the website. The official Epitomics website is as follows:
http://www.epitomics.com/products/product_info/57
As you can see, this...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (PBMC)
Specification PBMC
Preparation Cell harvesting/tissue preparation method: Ficoll-Hypaque, RBC lysis
Sample buffer: 2% human serum, 0.5 mM EDTA in PBS
Fixation Paraformaldehyde
Permeabilization No
Gating Strategy lymphocytes
Username

Abcam user community

Verified customer

Submitted Aug 03 2012

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