Product Description
FLT3 (D835Y) (Fms-like tyrosine kinase 3 with aspartic acid-to-tyrosine substitution at position 835), also known as a constitutively activating mutation in the tyrosine kinase domain, is a member of the receptor tyrosine kinase (RTK) family (subclass III) that plays a pivotal role in hematopoietic progenitor cell proliferation, differentiation, and survival. Wild-type FLT3 binds with high affinity to its ligand FLT3L (FLT3 ligand), leading to downstream activation of PI3K-AKT, RAS-MAPK, and STAT5 pathways. The D835Y mutation, located in the activation loop of the kinase domain, results in ligand-independent constitutive activation of FLT3, driving uncontrolled cell growth and survival. Beyond its role in normal hematopoiesis, dysregulation of FLT3 signaling, specifically the D835Y mutation, is closely associated with acute myeloid leukemia (AML), particularly in patients with poor prognosis and high relapse rates, as well as other hematological malignancies, making it a highly validated target for cancer therapy and drug discovery.
Screeningbio’s FLT3 (D835Y) Kinase Assay Kit provides a robust, sensitive, and high-throughput platform for quantifying FLT3 (D835Y) kinase activity and evaluating the potency of tyrosine kinase inhibitors (TKIs).
This kit utilizes the ADP-Glo Kinase Assay platform, a luminescent technology that measures the amount of ADP produced during the kinase reaction.
The assay is performed in two distinct steps:
Kinase Reaction: FLT3 (D835Y) catalyzes the transfer of a phosphate group from ATP to a specific substrate, resulting in the production of phosphorylated substrate and ADP.
ADP-Glo™ Reagent Addition: After the kinase reaction, the ADP-Glo™ Reagent is added to terminate the reaction and deplete any remaining unreacted ATP.
Kinase Detection: The Kinase Detection Reagent is added to simultaneously convert the produced ADP back into ATP, which is then used by luciferase to generate a luminescent signal.
The light intensity produced is directly proportional to the ADP concentration and, consequently, to the enzymatic activity of the FLT3 (D835Y) kinase.
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