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Human Amyloid Precursor Protein ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Amyloid Precursor Protein in Cerebral Spinal Fluid, Urine, Cell culture extracts, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.

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Images

Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (AB216944), expandable thumbnail
  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (AB216944), expandable thumbnail
  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (AB216944), expandable thumbnail
  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (AB216944), expandable thumbnail
  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (AB216944), expandable thumbnail

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Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Urine, Cell culture extracts, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

93.75 - 6000 pg/mL

Assay time

1h 30m

Sensitivity

= 14.9 pg/mL

Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info
-
Notes

-

Associated Products

Select an associated product type

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 synapses 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.

Alternative names

What's included?

1 x 96 Tests
Components
10X Human Amyloid Precursor Protein Capture Antibody
1 x 600 µL
10X Human Amyloid Precursor Protein Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
50X Cell Extraction Enhancer Solution (ab193971)
1 x 1 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 10 mL
Antibody Diluent 4BI
1 x 6 mL
Human Amyloid Precursor Protein Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 50 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human Amyloid Precursor Protein ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Amyloid Precursor Protein in Cerebral Spinal Fluid, Urine, Cell culture extracts, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma samples.

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Urine, Cell culture extracts, Tissue Extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

93.75 - 6000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 14.9 pg/mL

Precision

Intra assay

Sample

Serum

n
3
mean
-
SD
-
C.V.

6.2

Inter assay

Sample

Serum

n
5
mean
-
SD
-
C.V.

5.1

Recovery

Sample specific recovery

Sample type

Cell culture supernatant

Average %

= 112

Range

107 - 117 %

Sample type

Serum

Average %

= 112

Range

111 - 114 %

Sample type

EDTA Plasma

Average %

= 109

Range

93 - 123 %

Sample type

Cerebral Spinal Fluid

Average %

= 101

Range

92 - 107 %

Sample type

Urine

Average %

= 93

Range

86 - 100 %

Sample type

Cell culture extracts

Average %

= 93

Range

90 - 97 %

Sample type

Tissue Extracts

Average %

= 112

Range

107 - 118 %

Sample type

Heparin Plasma

Average %

= 106

Range

104 - 109 %

Sample type

Citrate plasma

Average %

= 91

Range

85 - 105 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Human Amyloid Precursor Protein ELISA Kit (ab216944) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Amyloid Precursor Protein protein in cell culture extracts, cell culture supernatant, cerebral spinal fluid, cit plasma, edta plasma, hep plasma, serum, tissue extracts, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Amyloid Precursor Protein with 14.9 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Amyloid Precursor Protein (GeneCards: APP) is a multifunctional transmembrane protein that consists of a 682 amino acid (aa) long extracellular domain, a 24 aa long transmembrane segment, and a 47 aa long cytoplasmic domain. Alternative splicing generates multiple isoforms including the most prevalent APP695, APP751, and APP770. 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. Amyloid Precursor Protein functions as a cell surface receptor and performs physiological functions on the surface of neurons relevant to neurite growth, neuronal adhesion and axonogenesis. Amyloid Precursor Protein is involved in cell mobility and transcription regulation through protein-protein interactions.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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.

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7 product images

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Example of human Amyloid Precursor Protein standard curve in Sample Diluent NS.

    Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Example of human Amyloid Precursor Protein standard curve in 1X Cell Extraction Buffer PTR.

    Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Interpolated concentrations of native Amyloid Precursor Protein in human serum and plasma samples.

    The concentrations of Amyloid Precursor Protein were measured in duplicates, interpolated from the Amyloid Precursor Protein standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 10%, plasma (citrate) 10%, plasma (heparin) 20% and plasma (EDTA) 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Amyloid Precursor Protein concentration was determined to be 52,942 pg/mL in neat serum, 63,882 pg/mL in neat plasma (citrate), 31,342 pg/mL in neat plasma (heparin), and 36,605 pg/mL in neat plasma (EDTA).

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Interpolated concentrations of native Amyloid Precursor Protein in human extract samples.

    The concentrations of Amyloid Precursor Protein were measured in three different dilutions in duplicate and interpolated from the Amyloid Precursor Protein standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in ng of Amyloid Precursor Protein per mg of extract (mean +/- SD, n=3). Amyloid Precursor Protein concentration was determined to be 213 ng/mg brain tissue extract, 8.24 ng/mg in liver tissue extract, 59.4 ng/mg in SH-SY5Y cell extract, 56.4 ng/mg in U-87 MG cell extract and 50.7 ng/mg in HeLa cell extract samples.

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Serum from nine individual healthy human male donors was measured in duplicate.

    Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Amyloid Precursor Protein concentration was determined to be 32,874 pg/mL with a range of 22,456 –‑ 42,097 pg/mL.

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Interpolated concentrations of native Amyloid Precursor Protein in human brain tissue extract, liver tissue extract, SH-SY5Y cell extract, HeLa cell extract and U-87 MG cell extract.

    Interpolated concentrations of native Amyloid Precursor Protein in human brain tissue extract based on a 20 μg/mL extract load, liver tissue extract based on a 500 μg/mL extract load, SH-SY5Y cell extract based on a 100 μg/mL extract load, HeLa cell extract based on a 25 μg/mL extract load, and U-87 MG cell extract based on a 100 μg/mL extract load. The concentrations of Amyloid Precursor Protein were measured in duplicate and interpolated from the Amyloid Precursor Protein standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Amyloid Precursor Protein concentration was determined to be 4,242 pg/mL in brain tissue extract, 4,224 pg/mL in liver tissue extract, 6,041 pg/mL in SH-SY5Y cell extract, 1,207 pg/mL in HeLa cell extract, and 5,676 pg/mL in U-87 MG cell extract.

  • Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944), expandable thumbnail

    Sandwich ELISA - Human Amyloid Precursor Protein ELISA Kit (ab216944)

    Interpolated concentrations of native Amyloid Precursor Protein in human cerebrospinal fluid (CSF), urine, A-549 cell culture supernatant, and HeLa cell culture supernatant (3 days).

    The concentrations of Amyloid Precursor Protein were measured in duplicates, interpolated from the Amyloid Precursor Protein standard curves and corrected for sample dilution. Undiluted samples are as follows: A-549 cell culture supernatant 25%, HeLa cell culture supernatant 25%, cerebrospinal fluid 0.5%, and urine 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Amyloid Precursor Protein concentration was determined to be 23,336 pg/mL in neat A-549 cell culture supernatant, 7,133 pg/mL in neat HeLa cell culture supernatant, 58,7958 pg/mL in neat cerebrospinal fluid, and 9,188 pg/mL in neat urine.

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