Recombinant human MLK1 (1–433) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. MLK1 or Mixed-Lineage Kinase 1 is a mitogen-activated protein kinase kinase kinase capable of activating the c-Jun NH(2)-terminal kinase (JNK) pathway. The catalytic domain of MLK1 has amino acid sequence similarity to both the tyr-specific and the ser/thr-specific kinase classes. In addition, MLK1 contains 2 leu/ile-zipper motifs and a basic sequence near its C-termini (1). MLK1 is threonine (and possibly serine) phosphorylated at multiple sites in the activation loop, with phosphorylation of threonine 312 required for full activation (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.Dorow, D.S. et al. (1993) Identification of a new family of human epithelial protein kinases containing two leucine/isoleucine-zipper domains. Europ. J. Biochem. 213, 701–10.
2.Durkin, J.T. et al. (2004) Phosphoregulation of mixed-lineage kinase 1 activity by multiple phosphorylation in the activation loop. Biochemistry 43, 16348–55.