EML4-ALK Kinase Enzyme System
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. 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.