Recombinant human MLK4 (amino acids 114-420) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. MLK4 or mixed lineage kinase 4 belongs to the superfamily of MAP kinase kinase kinases (MAP3K1) which contain both ser/thr and tyr kinases activity in their catalytic domains. The structure of this kinase family incorporates an N-terminal Src homology (SH3) domain, followed by the kinase domain, a leucine zipper region, and a CDC42 /RAC -interactive binding (CRIB) motif, but divergent C-terminal regions. MLK4 is highly expressed in kidney and pancreas. MLK4 is a negative regulator of TLR4. 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.
