The Kinase Enzyme Systems include a recombinant kinase enzyme, a substrate appropriate for the enzyme, a reaction buffer and supplemental reagents as needed. The FLT3 (ITD-NPOS) Kinase Enzyme System contains:
- FLT3 (ITD-NPOS) Kinase, 10μg (Human, recombinant; amino acids 571–993). MW: ~82kDa
- Native Swine Myelin Basic Protein (MBP) Substrate, (1mg/ml)
- 5X Reaction Buffer, 0.1M DTT
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”.
Application Note
NCBI Database Entry
The FLT3 (ITD-NPOS) 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.