ADP-Glo Kinase Assay + CDK2/CyclinE1 Kinase Enzyme System
Recombinant full-length human CDK2 and CyclinE1 were co-expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag on both proteins. CDK2 is a member of the Cyclin-Dependent Kinase family that is ubiquitously expressed. CDK2 is a catalytic subunit of the cyclin-dependent protein kinase complex, whose activity is restricted to the G1-S phase and essential for cell cycle G1/S phase transition. CDK2 associates with and is regulated by the regulatory subunits of the complex including Cyclin A or E, CDK inhibitor p21Cip1 (CDKN1A) and p27Kip1 (CDKN1B) (1). CDK2 phosphorylates multiple cellular substrates including SMAD3 and FOXO1. Phosphorylation of FOXO1 leads to its inhibition (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: CDK2/CyclinE1, 10ug (Human, recombinant full-length). MW: ~58kDa (CDK2) and ~73kDa (CyclinE1). Substrate: Native Histone H1 Protein; purified from calf thymus tissues. Other: Reaction Buffer, DTT. CDK2/CyclinE1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/1017/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.