Recombinant full-length human PKCθ was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Protein Kinase C theta (PKCθ) is an important component in the intracellular signaling cascade (1). Recent studies have suggested that local accumulation of fat metabolites inside skeletal muscle may activate a serine kinase cascade involving PKCθ leading to defects in insulin signaling and glucose transport in skeletal muscle (2). Insulin resistance plays a primary role in the development of type 2 diabetes and may be related to alterations in fat metabolism. PKCθ is a crucial component mediating fat-induced insulin resistance in skeletal muscle and is a potential therapeutic target for the treatment of type 2 diabetes (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: PKCθ, 10μg (Human, recombinant full-length). MW: ~110kDa. Substrate: PKCtide (ERMRPRKRQGSVRRRV); derived from protein kinase C epsilon (amino acids 149–164). Other: Reaction Buffer, DTT, Lipid Solution.
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.Manicassamy, S. and Sun, Z. (2007) The critical role of protein kinase C-theta in Fas/Fas ligand-mediated apoptosis. J. Immunol. 178, 312–9.
2.Kim, J.K. et al. (2004) PKC-theta knockout mice are protected from fat-induced insulin resistance. J. Clin. Invest. 114, 823–7.
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