Product Description
ABL(Y253F) is a key resistance mutation located in the P‑loop region of the ABL1 kinase domain. ABL1 is a member of the non‑receptor tyrosine kinase family that plays essential roles in cell proliferation, differentiation, adhesion, and apoptosis. In the context of the BCR‑ABL1 fusion oncoprotein—the primary driver of chronic myeloid leukemia (CML) and Philadelphia chromosome‑positive acute lymphoblastic leukemia (Ph+ ALL)—the Y253F mutation disrupts the conformation of the ATP‑binding loop, reducing the binding affinity of ATP‑competitive inhibitors such as imatinib while preserving aberrant kinase activity. This mutation frequently confers resistance to first‑ and second‑generation tyrosine kinase inhibitors and is closely associated with disease progression and poor clinical outcomes, making it a critical target for next‑generation inhibitor development and strategies to overcome drug resistance.
Screeningbio’s ABL(Y253F) Kinase Assay Kit provides a robust, sensitive, and high-throughput platform for quantifying ABL(Y253F) 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: ABL(Y253F) 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 ABL(Y253F) kinase.
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