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Alkaline Phosphatase Assay Kit (Colorimetric) ab83369 is a highly sensitive, simple, direct and HTS-ready colorimetric assay designed to measure alkaline phosphatase (ALP) activity in serum and other mammalian samples.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

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Images

Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (AB83369), expandable thumbnail
  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (AB83369), expandable thumbnail
  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (AB83369), expandable thumbnail
  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (AB83369), expandable thumbnail
  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (AB83369), expandable thumbnail

Publications

Key facts

Detection method

Colorimetric

Sample types

Plasma, Tissue Extracts, Cell culture media, Serum, Other biological fluids, Cell Lysate

Assay type

Enzyme activity (quantitative)

Reactive species

Mammals

Range

10 - 250 µU

Assay time

1h 10m

Sensitivity

> 10 µU

Associated Products

Select an associated product type

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Target data

Function

This isozyme plays a key role in skeletal mineralization by regulating levels of diphosphate (PPi).

Alternative names

What's included?

10000 Test
Components
ALP Assay Buffer I
20 x 100 mL
Enzyme Mix II
20 x 1 Vial
Stop Solution I
20 x 10 mL
pNPP Substrate
20 x 1 Vial

Recommended products

Alkaline Phosphatase Assay Kit (Colorimetric) ab83369 is a highly sensitive, simple, direct and HTS-ready colorimetric assay designed to measure alkaline phosphatase (ALP) activity in serum and other mammalian samples.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

Alternative names

Key facts

Detection method

Colorimetric

Sample types

Plasma, Tissue Extracts, Cell culture media, Serum, Other biological fluids, Cell Lysate

Assay type

Enzyme activity (quantitative)

Reactive species

Mammals

Range

10 - 250 µU

Assay time

1h 10m

Assay Platform

Microplate reader

Sensitivity

> 10 µU

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Alkaline Phosphatase Assay Kit (Colorimetric) ab83369 is a highly sensitive, simple, direct and HTS-ready colorimetric assay designed to measure alkaline phosphatase (ALP) activity in serum and other mammalian samples.

The kit is a pNPP assay; it uses p-nitrophenyl phosphate (pNPP) as a phosphatase substrate which turns yellow (λmax= 405 nm) when dephosphorylated by ALP.

Alkaline phosphatase assay protocol summary:
- add samples and standards to wells
- add pNPP solution to sample wells (not to standards)
- add ALP enzyme solution to standard wells (not to samples)
- incubate for 60 min at room temp
- add stop solution
- analyze with microplate reader

Technical Note: Alkaline phosphatase is unlikely to be detectable in healthy urine but may be elevated in certain kidney, liver or bone disorders.

This product is manufactured by BioVision, an Abcam company and was previously called K412 Alkaline Phosphatase Activity Colorimetric Assay Kit. K412-500 is the same size as the 500 test size of ab83369.

This kit contains 10 substrate tablets providing convenience for multiple usages.

pNPP Assays

pNPP assays are a class of phosphatase assays using p-nitrophenyl phosphate (pNPP) as a phosphatase substrate. They can be used to measure alkaline phosphatase, neutral phosphatase, and acid phosphatase activity. These different types of phosphatases are active respectively in alkaline, neutral, and acid assay buffers. pNPP assays include Alkaline Phosphatase Assay Kit ab83369 and Acid Phosphatase Assay Kit Acid Phosphatase Assay Kit (Colorimetric) ab83367.

Supplementary info

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

Activity summary

Alkaline phosphatase (ALP) is an enzyme that catalyzes the hydrolysis of phosphate groups from various molecules including nucleotides and proteins. It is a homo-dimeric protein with each monomer having a mass of about 57 kDa. The enzyme is widely expressed across different tissues with high levels present in the liver bone kidney and placenta. ALP is also commonly referred to as ALP or alkaline phosphatase where it exists as various isoenzymes depending on the tissue of expression.

Biological function summary

Alkaline phosphatase is involved in essential processes such as dephosphorylation which impacts cellular communication and energy metabolism. The enzyme participates in bone mineralization by breaking down pyrophosphate an inhibitor of mineralization to facilitate calcium and phosphate deposition. It does not typically function as part of a larger complex but its activity is critical in maintaining normal physiological pH levels in various tissues. The enzyme's activity is often measured using an alkaline phosphatase assay or ALP assay valuable for assessing different condition states within the body.

Pathways

Alkaline phosphatase plays roles in bone formation and liver function pathways. It interacts with other proteins like collagen and osteocalcin in the bone mineralization pathway which is vital for healthy bone development and maintenance. Additionally in liver function ALP is linked to other enzymes like gamma-glutamyltransferase (GGT) which together help detoxify hematopoietic and other bodily functions. Alkaline phosphatase activity can influence these pathways significantly impacting health status and treatment outcomes.

Associated diseases and disorders

Abnormal alkaline phosphatase levels often link to conditions such as Paget's disease of bone and hepatobiliary dysfunction. In Paget's disease elevated ALP indicates increased bone turnover often with increased osteocalcin activity. Meanwhile in liver disease altered ALP levels may connect with gamma-glutamyltransferase during diagnostic analyses to assess liver dysfunction or biliary obstruction. Regular monitoring of ALP activity is important for diagnosing these conditions and evaluating treatment responses.

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8 product images

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail
    Image courtesy of Hermenean A et al. Sci Rep. 2017; 7: 16641. doi: 10.1038/s41598-017-16599-5

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    ALP activity in cells undergoing osteogenic differentiation.

    Hermenean A et al. (2017) measured alkaline phosphatase activity in cells cultured in CHT/GO (3D chitosan scaffolds enriched with graphene oxide) biomaterials for up to 28 days using ab83369.

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail
    Image courtesy of Alesutan I et al. Sci Rep. 2017; 7: 2059. doi: 10.1038/s41598-017-01882-2. Reproduced under the creative commons license http://creativecommons.org/licenses/by/4.0/.

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    Effects of mineralocorticoid receptor blockade on phosphate-induced vascular calcification in vitro.

    Alesutan I et al. (2017) measured alkaline phosphatase activity in vitro in primary human aortic smooth muscle cells using ab83369. Cells were treated with phosphate (pi) or without (Ctr, and with or without additional treatment of spironolactone (Spr) or 10 μM eplerenone (Epl).

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail
    Image from Lee YM. et al., Biomed Res Int. 2019 Feb 19;2019:5073085. Fig 6b doi: 10.1155/2019/5073085. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    Alkaline phosphatase activity measured in the study of osteoporosis.

    Lee YM et al. (2019) used alkaline phosphatase assay kit ab83369 to measure the serum levels of alkaline phosphatase as a marker of bone turnover. Rat groups include sham-operated (Sham), OVX-control (OVX), E2-treated OVX groups (E2), and LABE 500 mg/kg treated OVX groups (LABE) (N=10/group).

    Serum ALP levels in the OVX group were significantly increased compared to those in Sham group, and oral administration of LABE (500 mg/kg) significantly reduced the serum ALP levels as an osteoporosis-related serum marker.

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail
    Image from Nepelska M et al., PLoS One 7(12), fig 8c. Doi: 10.1371/journal.pone.0052869. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    Nepelska M et al. (2012) assayed alkaline phosphatase activity in Caco-2 cells after 48 hours treatment with PMA and Butyrate using Alkaline Phosphatase assay kit (ab83369).

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    ALP measured in cell culture supernatants showing activity (U) per L of tested sample. Samples were diluted 1-16 fold.

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    ALP measured in biological fluids showing activity (U) per L of tested sample. Samples were diluted 4-64 fold.

  • Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369), expandable thumbnail

    Functional Studies - Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

    Measurement of ALP activity using ab83369. a. ALP activity in fresh medium (80 μL, without culturing), 3 day old HeLa cell culture medium (80 μL) and human serum (80 μL, 1/10 dilution). b. ALP activity in HeLa cells. 5 x 104 HeLa cells were homogenized in 1 mL of Assay Buffer, diluted 1/10 in Assay Buffer, and 80 μL used for the measurement.

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Product protocols

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