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AB83371

Alkaline Phosphatase Assay Kit (Fluorometric)

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(48 Publications)

Alkaline Phosphatase Assay Kit ab83371 is an addition-only ALP activity assay with just one 30 min incubation step. Readout on any fluorometric plate reader (Ex/Em 360/440 nm).

- Cited in over 40 publications
-Simple and fast protocol with results in one hour
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

View Alternative Names

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

5 Images
Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)
  • FuncS

Lab

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)

Alkaline phosphatase measured in biological fluids showing quantity (mU) per microliter of tested sample

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)
  • FuncS

PubMed

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)

Effects of IL-17A and/or TNF-α on alkaline phosphatase activity are shown. hMSCs were plated at a density of 5 x 103 cell/cm2, cells were cultured in osteogenic medium for 14 days, in the absence (0) or presence of TNF-α 1 ng/ml and/or IL-17A 50 ng/ml. ALP activity was measured by fluorometry useing ab83371. Results were analyzed using the Wilcoxon test. ∗p < 0.05; ∗∗p < 0.005 vs. induction medium alone.

Osta B et al., Front Immunol., 5:425. Figure 2.; doi:10.3389/fimmu.2014.00425. (2014)

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)
  • FuncS

Lab

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)

Alkaline phosphatase measured in mouse tissue lysates showing activity (mU) per mg protein of tested sample

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)
  • FuncS

Supplier Data

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)

4-Methylumbelliferone (4-MU) standard curve.

Other - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)
  • Other

Supplier Data

Other - Alkaline Phosphatase Assay Kit (Fluorometric) (AB83371)

Representative image of Alkaline Phosphatase Assay Kit (Fluorometric) ab83371

Components shown from left to right :

- MUP Substrate

- Enzyme Mix II

- Stop Solution VII

- ALP Assay Buffer I

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 kit.

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

Key facts

Detection method

Fluorescent

Sample types

Saliva, Urine (UTI), Plasma, Tissue Extracts, Cell culture media, Serum, Other biological fluids, Cell Lysate

Assay type

Enzyme activity

Results type

Quantitative

Assay time

1h

Assay Platform

Microplate reader

Reactivity data

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Product details

Alkaline Phosphatase Assay Kit (Fluorometric) (ab83371) is a highly sensitive kit to detect alkaline phosphatase (ALP) activity in serum and other biological samples. In the alkaline phosphatase assay protocol, ALP cleaves the phosphate group of the non-fluorescent 4-Methylumbelliferyl phosphate disodium salt (MUP) substrate, resulting in an increased fluorescent signal (Ex/Em = 360nm/440nm) when the substrate is dephosphorylated.

The kit is an ultra sensitive, simple, direct and HTS-ready assay designed to measure ALP activity with a detection sensitivity ~1 μU, more sensitive than colorimetric assays.

Alkaline phosphatase assay protocol summary:
- add samples and standards to wells
- add reaction mix to wells
- add ALP enzyme solution to standard wells (not to samples)
- incubate for 30 min at room temp
- add stop solution
- analyze with microplate reader

Technical Note: ALP is unlikely to be detectable in healthy urine but may be elevated in the presence of infection.

Related alkaline phosphatase assay products
For a colorimetric alkaline phosphatase assay kit, we recommend ab83369. which is based on the p-nitrophenyl phosphate (pNPP) assay substrate. pNPP turns yellow (405 nm) when dephosphorylated by ALP as part of an ALP activity assay.

For a luminescent alkaline phosphatase assay, we recommend ab233466, which is based on the ability of alkaline phosphatase to remove the phosphate group from D-luciferin phosphate. This releases the luciferin to be processed by luciferase.

For alkaline phosphatase staining, we recommend Alkaline Phosphatase Staining Kit ab284936, which is based on the substrate, 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) substrate. BCIP is processed by alkaline phosphatase to form a blue colored intermediate. Nitro Blue Tetrazolium (NBT) then oxidizes the intermediate to form an insoluble, dark blue dimer, which can be viewed by microscopy.

This product is manufactured by BioVision, an Abcam company and was previously called K422 Alkaline Phosphatase Activity Fluorometric Assay Kit. K422-500 is the same size as the 500 test size of ab83371.

Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters in alkaline buffer and produces an organic radical and inorganic phosphate. The change in alkaline phosphatase level and activity is associated with many diseases in the liver and bones.

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

What's included?

{ "values": { "500Test": { "sellingSize": "500 Test", "publicAssetCode":"ab83371-500Test", "assetComponentDetails": [ { "size":"1 x 25 mL", "name":"Stop Solution VII", "number":"AB83371-CMP04", "productcode":"" }, { "size":"1 x 1 Vial", "name":"MUP Substrate", "number":"AB83371-CMP03", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Enzyme Mix II", "number":"AB83371-CMP01", "productcode":"AB134678" }, { "size":"1 x 100 mL", "name":"ALP Assay Buffer I", "number":"AB83371-CMP02", "productcode":"AB171729" } ] } } }

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
Please refer to protocols

Supplementary information

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

Alkaline phosphatase also known as ALP is an enzyme with significant biological roles. It mechanically functions by catalyzing the hydrolysis of phosphate groups from various molecules including nucleotides proteins and alkaloids under alkaline conditions. ALP is a metalloenzyme possessing zinc and magnesium ions at its active site to aid in the reaction. This enzyme has a molecular mass of approximately 86 kDa and exists in dimeric form. ALP is expressed in many tissues with high levels in liver bone kidney and the placenta.
Biological function summary

ALP plays an important role in maintaining mineral balance and bone metabolism by breaking down pyrophosphate an inhibitor of mineralization. It facilitates the regulation of phosphate metabolism by contributing to phosphorus and calcium homeostasis. ALP is not part of a larger complex but interacts with calcium and phosphate ions to aid in the formation and resorption of bone mineral tissue. It serves as a marker in bone growth and liver function due to its prominent activity in those tissues visible in assays like the ALP staining method used to gauge enzyme presence and action.

Pathways

Alkaline phosphatase activity plays an essential role in the metabolic pathways related to bone mineralization and liver function. In bone tissue the enzyme participates in the calcification process important for bone strength and development by regulating inorganic phosphate availability. The liver expression of ALP is involved in bile excretion pathways where it dephosphorylates bile acids aiding in their conversion and recycling. It has a significant relationship with proteins such as osteocalcin in the bone mineralization pathway and bile acid transporters in the liver.

Alkaline phosphatase levels serve as diagnostic indicators for bone and liver diseases. High levels of ALP are often associated with bone disorders like Paget's disease and liver conditions like cholestasis. In these disorders ALP activity rises due to increased enzyme synthesis by osteoblasts in bone diseases or impaired bile flow in liver diseases. Furthermore the liver enzyme gamma-glutamyl transferase (GGT) is often measured alongside ALP to differentiate the cause of elevated ALP levels particularly to assess liver pathology.

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 (48)

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Magnesium-enhanced porcine particles using hydrothermal technique improve the osteogenic differentiation of cells.

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Low Magnesium Concentration Enforces Bone Calcium Deposition Irrespective of 1,25-Dihydroxyvitamin D Concentration.

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Usman Rashid,Sandra K Becker,Gerhard Sponder,Susanne Trappe,Mansur A Sandhu,Jörg R Aschenbach

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Repairing a critical cranial defect using WISP1-pretreated chondrocyte scaffolds.

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2022

Plasma rich in growth factors (PRGF) and leukocyte-platelet rich fibrin (L-PRF): comparative release of growth factors and biological effect on osteoblasts.

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Laura Baca-Gonzalez,Rebeca Serrano Zamora,Lisa Rancan,Francisco González Fernández-Tresguerres,Isabel Fernández-Tresguerres,Rosa M López-Pintor,Juan López-Quiles,Isabel Leco,Jesús Torres

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All-trans retinoic acid enhances the anti-tumour effects of fimaporfin-based photodynamic therapy.

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Judith Jing Wen Wong,Susanne Lorenz,Pål Kristian Selbo
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