5'-Nucleotidase Activity Assay Kit (Colorimetric) (BN01150)

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SKU:
BN01150
$825.63

Description

ELISA Kit Technical ManualMSDS

5'-Nucleotidase Activity Assay Kit (Colorimetric)

5'-Nucleotidase (5'-NT), also known at CD73 (EC 3.1.3.5) 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. Assay Genie'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.

Figure: (a) NH4+ standard curve (b) 5’-Nucleotidase specific activity in J774.2 mouse macrophage (60 µg protein) cell line lysate and rat liver tissue lysate (40 µg protein). Assays were performed following kit protocols.

Key Info Description

Product SKU

BN01150

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.

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