EPHB4 Kinase Enzyme System

Recombinant human EPHB4 (amino acids 561-end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. EPHB4 is a member of the Eph family of receptor tyrosine kinases which play a role in regulating cell adhesion and cell movement during embryonic development. Furthermore, because tissue disorganization and abnormal cell adhesion, movement, and survival characterize the more advanced stages of cancer, the inappropriate functioning of Eph receptor could play a causal role in malignancy. EPHB4 is uniquely expressed in the vascular endothelial and endocardial cells and animal knockout studies with EPHB4 reveal a phenotype that is similar to Ephrin B2 gene knockdown. 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.