ADP-Glo Kinase Assay + PKCμ Kinase Enzyme System

The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The PKCθ Kinase Enzyme System contains:

• PKCθ Kinase, 10μg (Human, recombinant full-length). MW: ~110kDa.
• PKCtide (ERMRPRKRQGSVRRRV) Substrate; derived from protein kinase C epsilon (amino acids 149164).
• Reaction Buffer, DTT, Lipid Solution.

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. 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. 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.

PKCθ NCBI Database Entry.

The PKCθ Kinase Enzyme System can be purchased with or without the ADP-Glo Kinase Assay reagents. Used together, the ADP-Glo Kinase Assay + Kinase Enzyme Systems provide a convenient method for profiling the effect of lead compounds on kinase activity. Assay advantages include broad dynamic range, ease of use and high sensitivity. Kinase Enzyme Systems are manufactured by SignalChem. Bulk quantities available upon request.

Use with ADP-Glo Kinase Assay

The 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 of chemical compounds 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.

See all Kinase Enzyme Systems available from Promega.