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5’-Nucleotidase (CD73) Activity Assay Kit (Colorimetric)

Simple, sensitive, enzymatic assay to measure 5'NT (CD73) Activity in biological samples

WARNING: This product can expose you to chemicals including Nickel Chloride, which is [are] known to the State of California to cause cancer.  For more information go to
Catalog #: K992

Product Details

Cat # +Size K992-100
Size 100 assays
Detection Method Colorimetric (OD 670 nm)
Species Reactivity Mammalian
Applications • Measurement of 5’-Nucleotidase activity in various tissues/cells
Features & Benefits • Simple, highly sensitive, high-throughput compatible
Kit Components • 5'-NT Assay Buffer
• 5'-NT Substrate
• 5'-NT Convertor
• 5'-NT Inhibitor
• 5'-NT Stop Solution
• 5'-NT Developer I
• 5'-NT Developer II
• NH4+ Standard (100 mM)
• 5'-NT Positive Control
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


5’-Nucleotidase (5’-NT), also known at CD73 (EC 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. BioVision’s 5’-Nucleotidase Activity 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 670 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 positive control

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Is this kit specific to CD73? Will it cross react with other enzymes such as CD39?
The substrate used in the kit is specific for CD73 and will not cross-react with CD39. The reaction with the substrate releases ammonia, which is then quantified and enzymes like alkaline phosphatase can also contribute to the generation of the final signal. Hence, we recommend the use of 5’-NT inhibitor to completely inhibit 5’-nucleotidase and distinguish from the signals generated by non-specific enzymes. Thus, the initial reaction is highly specific to CD73 due to substrate specificity.