Enterokinase Activity Fluorometric Assay Kit

• Cat.No.: Kit-1106
• Product Overview: Enterokinase or enteropeptidase is a heterodimeric serine protease produced by cells in the duodenal wall. It converts trypsinogen to trypsin, indirectly activating a number of pancreatic digestive enzymes. Enterokinase is a key enzyme for intestinal digestion of proteins. Its deficiency may cause severe protein malabsorption leading to impaired development and growth. Enterokinase also plays an important role in acute experimental and clinical pancreatitis. The high degree of specificity exhibited by enteropeptidase for cleaving affinity or other tags from recombinant proteins makes it an ideal tool for use in biochemical applications. Enterokinase Assay Kit employs a novel internally quenched 5-FAM/QXL FRET substrate for the detection of enterokinase activity. The enzyme cleaves the FRET substrate into two separate fragments resulting in the release of 5-FAM fluorescence which can be monitored at excitation /emission= 490/520 nm. The long wavelength fluorescence of 5-FAM is less interfered by the autofluorescence of components in biological samples and test compounds. The assays are performed in a convenient 96-well microplate format.

Specification

• Size: 100 Assays (96-well plate)
• Storage: Store all kit components at -20C, except for Component C; Store Component C at -80C; Protect Components A and B from light and moisture; Component D can be stored at room temperature for convenience.
• Shipping: Dry ice
• Kit Components: Component A: 5-FAM/QXL-520 Enterokinase substrate, Ex/Em=490/520 nm upon cleavage, 2 mM, 50 μL Component B: 5-FAM, fluorescence reference standard, Ex/Em=490/520 nm, 2 mM, 10 μL Component C: Recombinant Bovine Enterokinase, 40 μL Component D: 2X Assay Buffer, 25 mL Component E: Inhibitor E-64, 50 mM, 20 μL
• Materials Required but Not Supplied: 96-well microplate: Black, flat-bottom, 96-well plate with non-binding surface. Fluorescence microplate reader: Capable of detecting emission at 520 nm with excitation at 490 nm.