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Anti-Amyloid Precursor Protein antibody [Y188] ab32136 is a rabbit monoclonal antibody that is used in Amyloid Precursor Protein western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone Y188 has been tried and trusted by researchers since 2006 and is cited in >290 publications
- Specificity confirmed with APP knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

New 20 ul size available


Images

Immunoprecipitation - Anti-Amyloid Precursor Protein antibody [Y188] (AB32136), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Amyloid Precursor Protein antibody [Y188] (AB32136), expandable thumbnail
  • Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (AB32136), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Amyloid Precursor Protein antibody [Y188] (AB32136), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (AB32136), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPFlow Cyt (Intra)Flow CytWBICC/IF
Human
Tested
Tested
Not recommended
Not recommended
Tested
Tested
Mouse
Expected
Expected
Not recommended
Not recommended
Tested
Expected
Rat
Expected
Expected
Not recommended
Not recommended
Tested
Expected

Tested
Tested

Species

Human

Dilution info

1/500

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Rat, Human, Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/20000

Notes

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

Species

Rat

Dilution info

1/20000

Notes

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

Species

Human

Dilution info

1/20000

Notes

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

Tested
Tested

Species

Human

Dilution info

1/100 - 1/500

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

9 products for Alternative Product

1 product for Alternative Version

Target data

Function

Functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Interaction between APP molecules on neighboring cells promotes synaptogenesis (PubMed:25122912). 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 (By similarity). By acting as a kinesin I membrane receptor, plays a role in axonal anterograde transport of cargo towards synapes in axons (PubMed:17062754, PubMed:23011729). 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.Amyloid-beta 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. Amyloid-beta protein 42 is a more effective reductant than amyloid-beta protein 40. Amyloid-beta peptides bind to lipoproteins and apolipoproteins E and J in the CSF and to HDL particles in plasma, inhibiting metal-catalyzed oxidation of lipoproteins. APP42-beta 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).

Alternative names

Amyloid-beta precursor protein, APP, ABPP, APPI, Alzheimer disease amyloid protein, Amyloid precursor protein, Amyloid-beta A4 protein, Cerebral vascular amyloid peptide, PreA4, Protease nexin-II, CVAP, PN-II, APP, A4, AD1

Recommended products

Anti-Amyloid Precursor Protein antibody [Y188] ab32136 is a rabbit monoclonal antibody that is used in Amyloid Precursor Protein western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone Y188 has been tried and trusted by researchers since 2006 and is cited in >290 publications
- Specificity confirmed with APP knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

Y188

Purification technique

Affinity purification Protein A

Specificity

The antibody detects the C-terminal fragment of APP (APP-CTF).

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

The immunogen used for this product is within Human Amyloid Precursor Protein aa 750 to the C-terminus and therefore may detect gamma secretase fragments 50, 57 and 59 in addition to fragments C31, C80, C83 and C99. Cross-reactivity with these fragments has not been confirmed experimentally.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Amyloid Precursor Protein (APP) also known as amyloid protein is a transmembrane protein that is approximately 695 to 770 amino acids in length depending on the isoform. The molecular mass of APP can vary but typically falls around 100 to 140 kDa. It is heavily expressed in the central nervous system particularly in neurons but also in other tissues like muscle and kidney. The APP undergoes proteolytic processing which leads to the generation of various fragments including beta-amyloid peptides.

Biological function summary

The processing of APP plays a fundamental role in neuronal growth survival and repair. APP is cleaved into fragments that can regulate synaptic function and plasticity. It does not operate as a part of a complex but interacts with various cellular components. The protein participates in signaling pathways influencing cellular adhesion motility and neurite outgrowth. APP’s numerous interaction partners facilitate its involvement in different cellular processes highlighting its critical role in normal cell function.

Pathways

The APP is a central component in the amyloidogenic pathway where its cleavage by beta-secretase and gamma-secretase yields beta-amyloid. This pathway is one of two primary metabolic routes for APP—alternative enzymatic processing through the non-amyloidogenic pathway precludes beta-amyloid formation releasing peptides that do not aggregate. Enzymes like BACE1 (beta-secretase 1) and presenilin are important in the amyloidogenic pathway directly resulting in the production of the neurotoxic amyloid beta-peptide.

Associated diseases and disorders

APP is intensely linked to Alzheimer's disease and cerebral amyloid angiopathy. Accumulation of beta-amyloid peptides formed from APP cleavage is a hallmark of Alzheimer's disease leading to plaque formation in the brain. This aggregation impacts neuronal function and is associated with neurodegenerative processes. Interactions with proteins like tau are significant as tau also plays an essential role in Alzheimer's disease pathology. Misprocessing of APP and the resulting beta-amyloid aggregates are also contributors to cerebral amyloid angiopathy where deposits within cerebrovascular walls compromise vascular integrity.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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12 product images

  • Immunoprecipitation - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

    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.

    All lanes: Immunoprecipitation - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136)

    Predicted band size: 86 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

    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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and Alexa Fluor® 594 goat anti-mouse at a dilution of 1/500 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) . The secondary antibody was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody (ab15007) were used.

  • Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

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

    Lane 1: HeLa cell lysate at 20 µg

    Lane 2: Human fetal brain tissue lysate at 20 µg

    Lane 3: HEK293 cell lysate at 20 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 86 kDa

    Observed band size: 95 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

    ab32136 staining Amyloid Precursor Protein in wild-type HEK293 cells (top panel) and APP knockout HEK293 cells (Human APP (Amyloid Precursor Protein) knockout HEK-293T cell line ab255362) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32136 at 1/500 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136)

    Immunohistochemical staining of paraffin embedded human gliocytoma with purified ab32136 at a working dilution of 1/500. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) 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.

  • Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

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

    Lane 1: Mouse brain tissue lysate at 20 µg

    Lane 2: Rat brain tissue lysate at 20 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 86 kDa

    Observed band size: 95 kDa

  • Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

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

    All lanes: Hela cell lysate

    Predicted band size: 86 kDa

    Observed band size: 110 kDa, 95 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136)

    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.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail
    This image is courtesy of an anonymous customer review

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

    Blocked with 5% milk

    All lanes: Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136) at 1/1000 dilution

    All lanes: Mouse spleen whole cell lysate at 100 µg

    Secondary

    All lanes: HRP-conjugated goat anti-rabbit polyclonal IgG at 1/4000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 86 kDa

    Observed band size: 125 kDa, 95 kDa

    Exposure time: 10min

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136)

    Immunohistochemical analysis of formalin fixed paraffin embedded human cerebral cortex labelling Amyloid Precursor Protein with ab32136 at a concentration of 0.025µg/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20mins.

    ab32136 anti-Amyloid Precursor Protein antibody [Y188] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

    Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136)

    Immunohistochemical analysis of formalin fixed paraffin embedded human cerebral cortex labelling Amyloid Precursor Protein with ab32136 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

    ab32136 anti-Amyloid Precursor Protein antibody [Y188] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

    Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

  • Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136), expandable thumbnail

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

    Western blot: Anti-APP antibody [Y188] (ab32136) staining at 1/20000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32136 was shown to bind specifically to APP. A band was observed at 80-130 kDa in wild-type HAP1 and HEK-293T cell lysates with no signal observed at this size in both APP knockout cell lines. To generate this image, wild-type and APP knockout HAP1 and HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    Lanes 1 - 7: Western blot - Anti-Amyloid Precursor Protein antibody [Y188] (ab32136) at 1/20000 dilution

    Lanes 1 - 7: Western blot - Anti-Amyloid Precursor Protein antibody [Y188] - BSA and Azide free (Anti-Amyloid Precursor Protein antibody [Y188] - BSA and Azide free ab256586)

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: APP knockout HAP1 cell lysate at 20 µg

    Lane 3: Wild-type HEK-293T cell lysate at 20 µg

    Lane 4: APP knockout HEK-293T cell lysate at 20 µg

    Lane 5: Human Hippocampus cell lysate at 20 µg

    Lane 6: U-87 MG cell lysate at 20 µg

    Lane 7: K562 cell lysate at 20 µg

    Performed under reducing conditions.

    Observed band size: 80-130 kDa

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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