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Encompassing Amino Acids: 2-658(end)
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: The kinase assay is performed using the ADP-Glo™ Kinase Assay kit (Promega; Cat# V9101) which quantifies the amount of ADP produced. The ADP-Glo™ Reagent is added to terminate the reaction and to deplete the remaining ATP. The Kinase Detection Reagent is then added to convert ADP to ATP and to measure the newly synthesized ATP using a luciferase reaction.Assay: kinase activity was measured using a Poly-Glu/Tyr(4:1) substrate. The reaction was initiated by mixing Increasing amounts of the protein kinase with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl2, 0.1 mg/ml BSA, 250 µM DTT, and 0.2 mg/ml Poly-Glu/Tyr. The reaction was terminated by addition of the ADP-Glo™ Reagent, and the Kinase Detection Reagent was added. Phosphorylation was measured by detection of luminescence. The blank was determined from a "no kinase" sample.
Format: Aqueous buffer solution
Formulation: 50 mM sodium phosphate, pH 7.0, 300 mM NaCl, 150 mM imidazole, 0.1 mM PMSF, 0.25 DTT, 25% glycerol.
Genbank: NM_000061
Purification: affinity chromatography
Purity: ≥70%
Storage Stability: At least 6 months at -80°C.
Supplied As: Aqueous buffer solution.
Tags: C-terminal His-tag
Uniprot: Q06187
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)