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AB254503

Human Alkaline Phosphatase ELISA Kit

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(1 Publication)

Human Alkaline Phosphatase ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Alkaline Phosphatase with a sensitivity of 73 ng/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

AP-TNAP, TNS-ALP, TNSALP, Alkaline phosphatase liver/bone/kidney isozyme, Phosphoamidase, Phosphocreatine phosphatase, ALPL

3 Images
Sandwich ELISA - Human Alkaline Phosphatase ELISA Kit (AB254503)
  • sELISA

Supplier Data

Sandwich ELISA - Human Alkaline Phosphatase ELISA Kit (AB254503)

Interpolated concentrations of native ALPL in human Saos-2 cell extracts based on a 300 μg/mL extract load.

The concentrations of ALPL were measured in duplicate and interpolated from the ALPL standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ALPL concentration was determined to be 59.73 ng/mL.

Sandwich ELISA - Human Alkaline Phosphatase ELISA Kit (AB254503)
  • sELISA

Supplier Data

Sandwich ELISA - Human Alkaline Phosphatase ELISA Kit (AB254503)

Example of human ALPL standard curve in 1X Cell Extraction Buffer PTR.

The ALPL standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human Alkaline Phosphatase ELISA Kit (AB254503)
  • sELISA

Supplier Data

Sandwich ELISA - Human Alkaline Phosphatase ELISA Kit (AB254503)

Representative image of Human Alkaline Phosphatase ELISA Kit (ab254503)

Components shown from left to right :

- 10X Wash Buffer PT (ab206977)

- 10X Human Alkaline Phosphatase ELISA Kit Capture Antibody

- 10X Human Alkaline Phosphatase Detector Antibody

- 10X Human Alkaline Phosphatase ELISA Kit Lyophilised Purified Protein (2 vials)

- Human Alkaline Phosphatase ELISA Kit 50X Cell Extraction Enhancer Solution (ab193971)

- Sample diluent NS

- TMB solution

- Stop Solution

- 5X Cell Extraction Buffer PTR (ab193970)

- Antibody Diluent 4BR

SimpleStep Pre-Coated 96-Well Microplate (ab206978) - shown behind bottles / vials.

Plate Seals - shown underneath bottles / vials.

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the ELISA kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 73 ng/mL

Range

1.09 - 70 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Human ALPL ELISA Kit (ab254503) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of ALPL protein in cell lysate. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human ALPL with 73 ng/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 (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

ALPL, also known as tissue-nonspecific alkaline phosphatase, catalyzes phosphate ester hydrolysis when in a basic environment. Deficiency in this protein leads to loss in mineralization of bone and teeth which can be fatal. ALPL is currently being used a prognostic marker for the treatment of metastatic castration-resistant prostate cancer.

REACH authorisation
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.

Precision

[ { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.5" }, { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 24, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.7" } ]

Recovery

[ { "sample": "Cell Lysate", "range": "81 - 95 %", "average": "= 86" } ]

What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab254503-1x96Tests", "assetComponentDetails": [ { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB254503-CMP05", "productcode":"" }, { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB254503-CMP06", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB254503-CMP11", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB254503-CMP08", "productcode":"" }, { "size":"1 x 12 mL", "name":"Sample Diluent NS (ab193972)", "number":"AB254503-CMP13", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Development Solution", "number":"AB254503-CMP07", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seals", "number":"AB254503-CMP10", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human Alkaline Phosphatase Capture Antibody", "number":"AB254503-CMP04", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human Alkaline Phosphatase Lyophilized Recombinant Protein", "number":"AB254503-CMP02", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent 4BR", "number":"AB254503-CMP01", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human Alkaline Phosphatase Detector Antibody", "number":"AB254503-CMP12", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB254503-CMP09", "productcode":"" } ] } } }

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Supplementary information

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

Alkaline Phosphatase Tissue Non-Specific also referred to as ALPL or alkaline phosphatase protein functions mechanically as a hydrolase enzyme. It catalyzes the dephosphorylation of molecules contributing to phosphate and mineral metabolism. This enzyme with a molecular mass of approximately 57 kDa exhibits expression in various tissues including liver bone and kidneys. Researchers can use techniques like alkaline phosphatase ELISA and immunohistochemistry to study its expression and localization.
Biological function summary

Alkaline phosphatase in tissues plays a pivotal role in bone mineralization and development. It functions to hydrolyze phosphate groups releasing inorganic phosphate necessary for bone and teeth formation. It forms part of a larger enzyme complex that interacts with extracellular substrates ensuring the continuous supply of phosphate ions. This enzyme in particular impacts cellular processes involving calcification and cellular differentiation.

Pathways

Alkaline phosphatase operates within phosphate metabolism and signaling pathways. Within the skeletal system it forms part of the regulatory mechanism for osteoblast activity and bone formation. It relates to proteins like osteopontin and bone sialoprotein which regulate the mineralization process. These pathways ensure the balance between phosphate ions and mineral deposition in bone tissue.

Alkaline phosphatase abnormalities correlate with hypophosphatasia and rickets. Hypophosphatasia results from mutations in the ALPL gene causing defective bone mineralization. In cases of rickets improper phosphate metabolism can occur due to dysfunctional enzyme activity. The imbalance in alkaline phosphatase activity also connects to disorders like hyperparathyroidism where calcium and phosphate homeostasis is disrupted involving proteins such as parathyroid hormone.

Product protocols

Target data

Alkaline phosphatase that metabolizes various phosphate compounds and plays a key role in skeletal mineralization and adaptive thermogenesis (PubMed : 12162492, PubMed : 23688511, PubMed : 25982064). Has broad substrate specificity and can hydrolyze a considerable variety of compounds : however, only a few substrates, such as diphosphate (inorganic pyrophosphate; PPi), pyridoxal 5'-phosphate (PLP) and N-phosphocreatine are natural substrates (PubMed : 12162492, PubMed : 2220817). Plays an essential role in skeletal and dental mineralization via its ability to hydrolyze extracellular diphosphate, a potent mineralization inhibitor, to phosphate : it thereby promotes hydroxyapatite crystal formation and increases inorganic phosphate concentration (PubMed : 23688511, PubMed : 25982064). Acts in a non-redundant manner with PHOSPHO1 in skeletal mineralization : while PHOSPHO1 mediates the initiation of hydroxyapatite crystallization in the matrix vesicles (MVs), ALPL/TNAP catalyzes the spread of hydroxyapatite crystallization in the extracellular matrix (By similarity). Also promotes dephosphorylation of osteopontin (SSP1), an inhibitor of hydroxyapatite crystallization in its phosphorylated state; it is however unclear whether ALPL/TNAP mediates SSP1 dephosphorylation via a direct or indirect manner (By similarity). Catalyzes dephosphorylation of PLP to pyridoxal (PL), the transportable form of vitamin B6, in order to provide a sufficient amount of PLP in the brain, an essential cofactor for enzymes catalyzing the synthesis of diverse neurotransmitters (PubMed : 20049532, PubMed : 2220817). Additionally, also able to mediate ATP degradation in a stepwise manner to adenosine, thereby regulating the availability of ligands for purinergic receptors (By similarity). Also capable of dephosphorylating microbial products, such as lipopolysaccharides (LPS) as well as other phosphorylated small-molecules, such as poly-inosine : cytosine (poly I : C) (PubMed : 28448526). Acts as a key regulator of adaptive thermogenesis as part of the futile creatine cycle : localizes to the mitochondria of thermogenic fat cells and acts by mediating hydrolysis of N-phosphocreatine to initiate a futile cycle of creatine dephosphorylation and phosphorylation (By similarity). During the futile creatine cycle, creatine and N-phosphocreatine are in a futile cycle, which dissipates the high energy charge of N-phosphocreatine as heat without performing any mechanical or chemical work (By similarity).
See full target information Alkaline phosphatase, tissue-nonspecific isozyme

Publications (1)

Recent publications for all applications. Explore the full list and refine your search

PloS one 17:e0276328 PubMed36548302

2022

Xianling Gubao attenuates high glucose-induced bone metabolism disorder in MG63 osteoblast-like cells.

Applications

Unspecified application

Species

Unspecified reactive species

Xinlong Chen,Yan Li,Zhongwen Zhang,Liping Chen,Yaqian Liu,Shuhong Huang,Xiaoqian Zhang
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