5’-Nucleotidase (CD73) Activity Assay Kit (Colorimetric) (ab235945)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell Lysate, Tissue Lysate
Overview
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Product name
5’-Nucleotidase (CD73) Activity Assay Kit (Colorimetric)
See all CD73 kits -
Detection method
Colorimetric -
Sample type
Cell Lysate, Tissue Lysate -
Assay type
Quantitative -
Range
2 nmol/well - 15 nmol/well -
Species reactivity
Predicted to work with: Mammals
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Product overview
5’-Nucleotidase (CD73) Activity Assay Kit (Colorimetric) (ab235945) is a simple two-step end-point assay for the measurement of 5’-Nucleotidase activity in various tissues/cells. This assay 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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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests 5’-NT Assay Buffer 1 x 25ml 5’-NT Convertor 1 vial 5’-NT Developer I 1 x 8ml 5’-NT Developer II 1 x 4ml 5’-NT Inhibitor 1 x 250µl 5’-NT Positive Control 1 vial 5’-NT Stop Solution 1 x 500µl 5’-NT Substrate 1 vial NH4+ Standard (100 mM) 1 x 100µl -
Research areas
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Function
Hydrolyzes extracellular nucleotides into membrane permeable nucleosides. -
Involvement in disease
Defects in NT5E are the cause of calcification of joints and arteries (CAJA) [MIM:211800]. A condition characterized by adult-onset calcification of the lower extremity arteries, including the iliac, femoral and tibial arteries, and hand and foot capsule joints. Age of onset has been reported as early as the second decade of life, usually involving intense joint pain or calcification in the hands. -
Sequence similarities
Belongs to the 5'-nucleotidase family. -
Cellular localization
Cell membrane. - Information by UniProt
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Alternative names
- 5' NT
- 5' nucleotidase (CD73)
- 5' nucleotidase precursor
see all
Images
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Example Data
NH4+ standard curve.
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Example Data5’-NT specific activity in J774.2 mouse macrophage (60 µg protein) cell line lysate and rat liver tissue lysate (40 µg protein).
