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AB138887

Alkaline Phosphatase Assay Kit (Fluorometric - Near Infrared)

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Alkaline Phosphatase Assay kit (Fluorometric-Near Infrared) (ab138887) uses our SunRed Dye based substrate.

View Alternative Names

ALPPL, ALPPL2, ALPG, ALP-1, Alkaline phosphatase Nagao isozyme, Germ cell alkaline phosphatase, Placental alkaline phosphatase-like, GCAP, PLAP-like

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Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric - Near Infrared) (AB138887)
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Supplier Data

Functional Studies - Alkaline Phosphatase Assay Kit (Fluorometric - Near Infrared) (AB138887)

Sample Standard Curve for Alkaline phosphatase. Alkaline phosphatase dose response was measured with the Alkaline Phosphatase Assay Kit (Fluorometric –Near Infrared) in a solid black 96-well plate using a Gemini microplate reader (Molecular Devices). As low as 0.3 mU/mL alkaline phosphatase can be detected with 60 minutes incubation (n=3).

Key facts

Detection method

Fluorescent

Sample types

Cell culture extracts, Cell culture supernatant

Assay type

Enzyme activity (quantitative)

Sensitivity

= 0.3 mU/mL

Assay Platform

Microplate reader

Reactivity data

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Product details

Alkaline Phosphatase Assay kit (Fluorometric-Near Infrared) (ab138887) uses our SunRed Dye based substrate. The weakly fluorescent SunRed Dye phosphate is sensitive to phosphatase-induced hydrolysis, giving the SunRed Dye fluorophore that possesses intense red fluorescence. Upon phosphatase-induced hydrolysis, the SunRed Dye phosphate solution has its absorption blue-shifted more than 100 nm. The maximum absorption of SunRed Dye fluorophore at 633 nm makes this substrate an ideal NIR probe that can be readily detected with many fluorescence instrument systems often equipped with Cy5 settings. Based on the near infrared fluorescence of SunRed Dye fluorophore, the signal can be easily read by a fluorescence microplate reader at Ex/Em = ~630/660 nm.

ab138887 is designed to accommodate high throughput screening of protein phosphatase inhibitors due to its low interference from biological samples. It can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.

Alkaline phosphatase is widely used in various biological assays (in particular, immunoassays) and ELISA-based diagnostics.

What's included?

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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
-20°C

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 can hydrolyze various phosphate compounds.
See full target information ALPG

Additional targets

ALPP,Alkaline phosphatase, tissue-nonspecific isozyme,ALPI
websiteProtocolBooklet
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