Anti-Amyloid Precursor Protein antibody [Y188] (ab32136)

Overview

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • PurityProtein A purified
  • ClonalityMonoclonal
  • Clone numberY188
  • IsotypeIgG
  • Research areas

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 1/20000. Detects a band of approximately 95 kDa (predicted molecular weight: 87 kDa).

For unpurified, use 1/100 - 1/10000.

IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/70.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/30.
ICC/IF 1/100.
IHC-FoFr 1/750. PubMed: 18974297
IHC-Fr Use at an assay dependent concentration.

Target

  • 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

Anti-Amyloid Precursor Protein antibody [Y188] images

  • Immunofluorescence staining of SH-SY5Y cells with purified ab32136 at a working dilution of 1 in 100, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab32136 was used at a dilution of 1/200 followed by an Alexa Fluor® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  • All lanes : Anti-Amyloid Precursor Protein antibody [Y188] (ab32136) at 1/20000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : Human fetal brain tissue lysate
    Lane 3 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 87 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-Amyloid Precursor Protein antibody [Y188] (ab32136) at 1/20000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 87 kDa
    Observed band size : 95 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human gliocytoma with purified ab32136 at a working dilution of 1/500. The secondary antibody used is ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • ab32136 (purified) at 1/30 immunoprecipitating amyloid beta precursor protein in A431 (Lane 1). Lane 2 - PBS. For western blotting a HRP-conjugated anti-rabbit IgG specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST. Diluting buffer and concentration: 5% NFDM /TBST.
  • Overlay histogram showing PC-12 cells fixed in 2% PFA and stained with purified ab32136 at a dilution of 1 in 70 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 150. Rabbit monoclonal IgG was used as an isotype control (black) and cells without antibody were used as a negative control (blue).
  • Anti-Amyloid Precursor Protein antibody [Y188] (ab32136) at 1/20000 dilution (unpurified) + 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.
  • Unpurified 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.

  • Unpurified 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 Precursor Protein antibody [Y188] (ab32136) at 1/1000 dilution (unpurifed) + 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

    Blocked with 5% milk

    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 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 18 Publications for this product

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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...

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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 ...

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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...

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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...

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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...

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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

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Verified customer

Submitted Aug 03 2012

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