The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The HGK Kinase Enzyme System contains:
- HGK Kinase, 10μg (Human, recombinant; amino acids 1–328). MW: ~64kDa
- Native Swine Myelin Basic Protein (MBP) Substrate (1mg/ml)
- 5X Reaction Buffer, 0.1M DTT
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-α signaling.
Application Note
NCBI Database Entry
The HGK Kinase Enzyme System can be purchased with or without the ADP-Glo™ Kinase Assay reagents. Used together, the ADP-Glo™ Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.
Use with ADP-Glo™ Kinase Assay
The 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 of chemical compounds 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.