ADP-Glo Kinase Assay + CDK6/CyclinD3 Kinase Enzyme System
Recombinant full-length human CDK6 and CyclinD3 were co-expressed by baculovirus in Sf9 insect cells using an N-terminal His tag on both proteins. CDK6 is a member of the cyclin-dependent family of protein kinases that are important regulators of cell cycle progression. CDK6 activity is regulated by the D-type cyclins and members of the INK4 family of CDK inhibitors (1). The CDK6 kinase activity is detected in mid-G1 phase of the cell cycle and is responsible for the phosphorylation and regulation of the activity of tumor suppressor protein Rb. Although CDK6 and CDK4 can both phosphorylate multiple residues in the Rb protein, they do so with different residue selectivities in vitro; CDK6 phosphorylates Thr821 while CDK4 phosphorylates Thr826 on Rb protein (2). 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 chemical compounds have 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. Kinase Enzyme System contains: Kinase: CDK6/CyclinD3, 10ug (Human, recombinant full-length). MW: ~40kDa (CDK6) and ~35kDa (CyclinD3). Substrate: Native Histone H1 Protein; purified from calf thymus tissues. Other: Reaction Buffer, DTT. CDK6/CyclinD3 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/1021/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.