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, 10μg (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.
Profile More Compounds In-House: ADP-Glo™ Kinase Assay + Kinase Enzyme System is optimized so that you are up and running in no time. Complete Systems: The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer, DTT and supplemental reagents as needed. Obtain Reliable Results: The broad dynamic range, the ease of use and better sensitivity obtained with ADP-Glo™ Kinase Assay result in less ambiguous data.
Notes
Kinase Enzyme System manufactured by SignalChem.
Bulk quantities available upon request.
References
1.Zhao, Z. et al. (2004) Rho-associated kinases play a role in endocardial cell differentiation and migration. Dev Biol. 275, 183–91.
2.Noma, K. et al. (2006) Physiological role of ROCKs in the cardiovascular system. Am J Physiol Cell Physiol. 290, C661–8.
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