Description
Convenient, Scalable Kinase Profiling
The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The InsR (R1201Q) Kinase Enzyme System contains:
InsR (R1201Q) Kinase, 10μg (Human, recombinant; amino acids 1011–end). MW: ~70kDa
AXLtide Substrate (1mg/ml) (KKSRGDYMTMQIG); derived from mouse Insulin receptor substrate 1 amino acids 979–989
5X Reaction Buffer, 2.5M MnCl 2 , 0.1M DTT
Recombinant human InsR (amino acids 1011–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. InsR is the insulin receptor tyrosine kinase that is involved in insulin signaling. InsR is post-translationally cleaved into two chains, α and β, that are covalently linked. Binding of insulin to the InsR stimulates glucose uptake. Insulin receptor signaling helps to maintain fuel homeostasis and prevent diabetes. Studies have shown that a conditional knockout of insulin receptor substrate 2 (IRS2) in mouse pancreas β cells and parts of the brain—including the hypothalamus—increased appetite, lean and fat body mass, linear growth, and insulin resistance that progressed to diabetes. InsR signaling also increases the regeneration of adult β cells and the central control of nutrient homeostasis.
The InsR (R1201Q) 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.
