ADP-Glo Kinase Assay + ZAK Kinase Enzyme System
Recombinant full-length human ZAK was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ZAK is a member of the MAPKKK family of signal transduction molecules and mediates gamma radiation signaling leading to cell cycle arrest. The activity of ZAK plays a role in cell cycle checkpoint regulation as well as being involved in regulating actin organization (1). Expression of kinase-dead ZAK in mouse fibroblasts disrupts actin stress fibers and causes morphologic changes. ZAK can activate JNK through MKK4/MKK7 and ERK5/p38-gamma via MKK3/MKK6. Expression of ZAK increases the population of cells in the G2/M phase of the cell cycle, whereas dominant-negative ZAK attenuated the G2 arrest caused by gamma radiation (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. Kinase Enzyme System contains: Kinase: ZAK, 10ug (Human, recombinant full-length). MW: ~82kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT. ZAK NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/51776/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.