Product Description
FGFR3 (K650E) (Fibroblast growth factor receptor 3 with lysine-to-glutamic acid substitution at position 650), also known as a constitutively activating mutation in the tyrosine kinase domain, is a member of the receptor tyrosine kinase (RTK) family that plays a pivotal role in bone development, chondrocyte proliferation and differentiation, and cell growth regulation. FGFR3 normally binds with high affinity to ligands such as FGF1, FGF2, FGF4, FGF8, and FGF9, leading to downstream signaling through RAS-MAPK and PI3K-AKT pathways. The K650E mutation results in ligand-independent, constitutive activation of FGFR3, driving uncontrolled downstream signaling and pathological cell proliferation. Beyond its role in skeletal development, dysregulation of FGFR3 signaling, specifically the K650E mutation, is closely associated with multiple cancers including bladder cancer (especially in non-muscle-invasive tumors), cervical cancer, multiple myeloma, and seminoma, as well as severe skeletal dysplasias such as thanatophoric dysplasia type II (a lethal neonatal dwarfism disorder), making this mutant a highly validated target for cancer therapy and drug discovery.
Screeningbio’s FGFR3 (K650E) Kinase Assay Kit provides a robust, sensitive, and high-throughput platform for quantifying FGFR3 (K650E) 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: FGFR3 (K650E) 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 FGFR3 (K650E) kinase.
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