ADP-Glo Kinase Assay + FLT3 Kinase Enzyme System
Recombinant human FLT3 (571-993) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. FLT3 is a receptor tyrosine kinase that has been shown to play a role in proliferation and survival of hematopoietic progenitor cells as well as differentiation of early B lymphoid progenitors (1). FLT3 consists of an extracellular domain composed of five immunoglobulin-like domains, one transmembrane region and a cytoplasmic kinase domain split into two parts by a kinase-insert domain. FLT3 is the most frequently mutated gene in cases of acute myelogenous leukemia (AML). About 30% to 35% of patients have either internal tandem duplications (ITDs) in the juxtamembrane domain or mutations in the activating loop of FLT3 (2). The consequence of either FLT3-ITD or activating loop mutations is the constitutive activation of the tyrosine kinase activity. 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: FLT3, 10ug (Human, recombinant; amino acids 571-993). MW: ~73kDa. Substrate: Native Swine Myelin Basic Protein (MBP). Other: Reaction Buffer, DTT. FLT3 NCBI Database Entry: www.ncbi.nlm.nih.gov/gene/2322/. Visit www.promega.com/kinase/ to see all Kinase Enzyme Systems.