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FAK Kinase Assay

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Compound Test Services

CT-001

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Product Description


FAK (Focal adhesion kinase), also known as PTK2 (protein tyrosine kinase 2), is a member of the non-receptor tyrosine kinase family that plays a pivotal role in integrin-mediated cell adhesion, migration, invasion, proliferation, and survival. FAK is activated by autophosphorylation at Tyr397 in response to integrin clustering, growth factor receptor signaling, or mechanical stimuli, leading to recruitment of Src family kinases and downstream activation of pathways such as PI3K-AKT and MAPK. Beyond its role in normal cell physiology and embryonic development, dysregulation of FAK signaling—through overexpression, hyperactivation, or gene amplification—is closely associated with tumor progression, metastasis, epithelial-to-mesenchymal transition (EMT), angiogenesis, and chemoresistance in multiple cancers including breast, colorectal, glioblastoma, and pancreatic cancer, making it a highly validated target for cancer therapy and drug discovery.


Screeningbio’s FAK Kinase Assay Kit provides a robust, sensitive, and high-throughput platform for quantifying FAK 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:

  1. Kinase Reaction: FAK catalyzes the transfer of a phosphate group from ATP to a specific substrate, resulting in the production of phosphorylated substrate and ADP.

  2. ADP-Glo™ Reagent Addition: After the kinase reaction, the ADP-Glo™ Reagent is added to terminate the reaction and deplete any remaining unreacted ATP.

  3. 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 FAK kinase.




Data

TKs Kinase Inhibition Assay. Staurosporine was titrated using established assay protocol. Non-linear regression was used to plot RLU signal vs. [Compound, M], and EC50 /IC50 values were determined, using GraphPad Prism software.
TKs Kinase Inhibition Assay. Staurosporine was titrated using established assay protocol. Non-linear regression was used to plot RLU signal vs. [Compound, M], and EC50 /IC50 values were determined, using GraphPad Prism software.


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