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 EML4-ALK Kinase Enzyme System contains:
EML4-ALK Kinase, 10μg [Human, recombinant; amino acids 1–496 (EML4) and 1058–end (ALK)]. MW: ~150kDa
IGF1Rtide Substrate (1mg/ml), (KKKSPGEYVNIEFG); derived from human IRS-1 protein residues 891–902
5X Reaction Buffer, 2.5M MnCl 2 , 0.1M DTT
Recombinant human fusion protein EML4 (amino acids 1–496)-ALK (amino acids 1058–end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. ALK or CD246 is a receptor tyrosine kinase, which belongs to the insulin receptor superfamily, which comprises an extracellular domain, a hydrophobic stretch corresponding to a single pass transmembrane region, and an intracellular kinase domain. CD246 has been found to be rearranged, mutated, or amplified in a series of tumours including anaplastic large cell lymphomas, neuroblastoma, and non-small cell lung cancer. EML4-ALK mutations in lung cancer confer resistance to ALK inhibitors.
The EML4-ALK 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.
