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Recombinant human FLT3 (ITD-NPOS) (571-993) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag. Fms-related tyrosine kinase 3 (FLT3) is a gene that encodes for a tyrosine kinase that activates pathways in hematopoietic cells important in cellular proliferation. FLT3 is frequently mutated in acute myeloid leukemia (AML), myelodysplastic syndromes, other hematologic malignancies, and colorectal cancer. FLT3 mutations are observed in 24.3% of AML, of which internal tandem duplications (ITD) are the most frequent. FLT3 ITD occurs when sequences of less than ten to several hundred bases in length are repeated within the juxtamembrane domain are always “in-frame”. 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.
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