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Recombinant human HGK (amino acids 1-328) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. HGK is the mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4) and a member of the serine/threonine protein kinase family. HGK has been shown to specifically activate MAPK8/JNK. The activation of MAPK8 by HGK can be inhibited by dominant-negative mutants of MAP3K7/TAK1, MAP2K4/MKK4, and MAP2K7/MKK7, which suggest that this kinase functions through the MAP3K7-MAP2K4-MAP2K7 kinase cascade and mediates TNF-alpha signaling. 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.
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