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.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Matsumoto, S. et al. (2004) Characterization of a MAPKK-like protein kinase TOPK. Biochem Biophys Res Commun. 325, 997–1004.
2.Simons-Evelyn, M. et al. (2001) PBK/TOPK is a novel mitotic kinase which is upregulated in Burkitt's lymphoma and other highly proliferative malignant cells. Blood Cells Mol Dis 27, 825–9.