General description
5′-Nucleotidase (5′-NT), also known at CD73 is an enzyme
located in the plasma membrane. It converts extracellular nucleotides like
5′-AMP to their corresponding nucleosides, through phosphorylitic cleavage.
This conversion facilitates uptake of the nucleosides through nucleoside
receptors into the cell, where they can again be phosphorylated to generate
nucleotides and contribute to the nucleotide pool, inside the cell. 5′-NT
levels are elevated in hepatic diseases such as viral hepatitis, alcoholic liver
disease and cirrhosis.
Features and Benefits
5′-Nucleotidase (CD73) Activity Assay Kit:
- Sensitive,
detecting as low as 0.2 mU of 5′-NT.
- Compatible
with high-throughput handling systems.
Suitability
The kit is suitable for the measurement of 5′-Nucleotidase
activity in animal tissue lysates (eg. liver), cell lysates, recombinant enzyme
and purified protein.
Principle
The 5′-Nucleotidase (CD73) Activity Assay Kit is a simple
two-step end point assay that relies on the Berthelot′s test for quantification
of ammonia. In this assay, the action of 5′-nucleotidase on the substrate
generates a product, which releases ammonia in presence of the converter.
Developer I and II are then used to quantify the released ammonia through
increase in absorbance at A670 nm. This assay can detect as low
as 0.2 mU of 5′-NT. Since non-specific enzymes like alkaline phosphatase can
give a positive signal in this assay, 5′-NT inhibitor may be used to completely
inhibit 5′-nucleotidase and distinguish from the signal from non-specific
enzymes. The assay kit also includes 5′- Nucleotidase (5′-NT) enzyme for use as
a positive control.
