ADP-Glo Kinase Assay + CDK9/CyclinK Kinase Enzyme System
Recombinant full-length human CDK9 and CyclinK proteins were co-expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. CDK9/CyclinK is a member of the cyclin-dependent protein kinase (CDK) family. CDK9 is closely related to cdc28 and cdc2 and is an important regulator of the cell cycle (1). CDK9 is a component of the multiprotein complex TAK/P-TEF beta. CDK9 can modulate RNA polymerase II-directed transcription by phosphorylating the C-terminal domain of the largest subunit of RNA polymerase II. CDK9 forms a complex with and is regulated by its regulatory subunit cyclin T or cyclin K. CDK9 also interacts with the HIV-1 Tat protein, which suggests a possible involvement of this protein in AIDS (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: CDK9/CyclinK, 10ug (Human, recombinant full-length). MW: ~68kDa (CDK9) and ~67kDa (CyclinK). Substrate: PDKtide (KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC); residues 1-14 are derived from AKT1 (307-320), and residues 16-39 are derived from PKN2/PRK2 (961-984). Other: Reaction Buffer, DTT. CDK9/CyclinK NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/1025/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.