ADP-Glo Kinase Assay + ROCK1 Kinase Enzyme System
Recombinant human ROCK1 (amino acids 17-535) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ROCK1 is a ubiquitously expressed serine-threonine kinase that is a downstream target of the small GTPase RhoA. ROCK1 is involved in diverse cellular functions, including smooth muscle contraction, actin cytoskeleton organization, cell adhesion and motility, and gene expression. ROCK1 contributes to the development of cardiac fibrosis and induction of fibrogenic cytokines in cardiomyocytes in response to pathological stimuli. ROCK1 knockout mice exhibit reduced perivascular and interstitial fibrosis, which is associated with reduced expression of a variety of extracellular matrix (ECM) proteins and fibrogenic cytokines. 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: ROCK1, 10ug (Human, recombinant; amino acids 17-535). MW: ~85kDa. Substrate: S6K substrate (KRRRLASLR); derived from human 40S ribosomal protein S6 (amino acid 230-238). Other: Reaction Buffer, DTT. ROCK1 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/6093/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.