ADP-Glo Kinase Assay + TOPK Kinase Enzyme System
Recombinant full-length human TOPK was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. TOPK is a MAPK kinase that phosphorylates p38 MAPK and is activated in a cell-cycle-dependent manner in neuronal progenitor cells in vitro (1). Expression of TOPK is detected in male germ line progenitor cells, activated T-cells and a variety of lymphomas and leukemias. In vitro studies have shown that activated TOPK phosphorylates p38 MAPK but not JNK or ERK. TOPK activation requires phosphorylation by both the M-phase CDK1/CyclinB kinase complex and another unknown kinase, possibly Raf C or Raf A. TOPK may play an important role in linking extracellular signals to an intracellular state, possibly allowing extracellular influence on the cell-cycle-related processes of proliferation or differentiation (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: TOPK, 10ug (Human, recombinant full-length). MW: ~68kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT. TOPK NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/55872/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.