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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: Kinase activity was measured using a Poly-Glu/Tyr(4:1) substrate. Increasing amounts of the protein kinase were mixed with 0.25 mg/ml substrate in 20 µl final volume. The blank was determined from a "no substrate" sample. The reaction was initiated by addition of 5 µl of [33P]-ATP buffer containing 10 mM ATP, 1 m Ci [33P]-ATP, 25 mM MOPS, 12.5 mM β-glycero-phosphate, 25 mM MgCl2, 2 mN EDTA, 0.25 mM fresh DTT, pH 7.2. After incubating for 30°C for 15 minutes, the reaction was terminated by spotting 20 µl of the mixture onto phosphocellulose paper strips that were fixed in 1% phosphoric acid. Radioactivity was determined using a scintillation counter.
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: ≥85%
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)