DNA-PK Kinase Enzyme System

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The DNA-PK Kinase Enzyme System contains:

• DNA-PK Kinase, 2,500 units (Human, native full-length). MW: ~460kDa (catalytic subunit); ~85kDa (Ku subunit 1); ~70kDa (Ku subunit 2). One unit is the amount of enzyme required to incorporate 1pmol of phosphate into DNA-PK Peptide Substrate (Cat.# V5671) in 1 minute at 30°C.
• Storage Buffer: 25mM HEPES (pH 7.5), 50mM KCl, 0.2mM EDTA, 10mM MgCl₂, 1mM DTT, 10% glycerol.
• DNA-Dependent Protein Kinase Peptide Substrate (EPPLSQEAFADLWKK).
• Reaction Buffer, DNA-PK Activation Buffer (100µg/ml calf thymus DNA in 1X TE.), DTT.

Native human DNA-Dependent Protein Kinase (DNA-PK) purified from HeLa cells. DNA-PK consists of an approximately 460kDa catalytic subunit and a heterodimeric DNA-binding subunit (Ku) containing an 85kDa and an 70kDa peptide. The gene sequence can be found at accession number NM_006904.

DNA-PK NCBI Database Entry.

The DNA-PK Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.

Use with ADP-Glo™ Kinase Assay

The ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is a substrate in a reaction catalyzed by Ultra-Glo™ Luciferase that produces light. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects of chemical compounds on the activity of a broad range of purified kinases, making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.

See all Kinase Enzyme Systems available from Promega.