ADP-Glo Kinase Assay + ROCK2 Kinase Enzyme System
Recombinant human ROCK2 (5-554) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ROCK2 is a ubiquitously expressed serine/threonine kinase localized in the nucleus that regulates cytokinesis, smooth muscle contraction, the formation of actin stress fibers and focal adhesions, and the activation of the c-fos serum response element (1). ROCK2 is an immediate downstream target of the small GTPase RhoA. ROCK2 may play a pivotal role in cardiovascular diseases such as vasospastic angina, ischemic stroke and heart failure. Inhibition of ROCKs by statins or other selective inhibitors leads to the upregulation and activation of endothelial nitric oxide synthase (eNOS) and reduction of vascular inflammation and atherosclerosis (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: ROCK2, 10ug (Human, recombinant; amino acids 5-554). MW: ~88kDa. Substrate: S6K Substrate (KRRRLASLR); derived from human 40S ribosomal protein S6 (amino acids 230-238). Other: Reaction Buffer, DTT. ROCK2 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/9475/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.