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CDC7/DBF4 Kinase Assay

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Compound Test Services

CT-001

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Product Description


CDC7/DBF4 is a serine/threonine kinase complex composed of the catalytic subunit CDC7 (cell division cycle 7) and its regulatory subunit DBF4 (Dumbell former 4), which plays a pivotal role in the initiation of DNA replication, cell cycle progression, and genome stability. The CDC7/DBF4 complex phosphorylates key substrates such as MCM proteins (minichromosome maintenance complex components), thereby activating replication origins and ensuring proper S-phase entry. Beyond its essential physiological functions in cell division, dysregulation of CDC7/DBF4 signaling—including overexpression, aberrant activation, and altered complex formation—is frequently observed in a wide range of human cancers, such as breast, ovarian, colorectal, and hematologic malignancies, contributing to uncontrolled proliferation, replication stress, and therapeutic resistance. These features have established CDC7/DBF4 as a highly validated target for cancer therapy and drug discovery.


Screeningbio’s CDC7/DBF4 Kinase Assay Kit provides a robust, sensitive, and high-throughput platform for quantifying CDC7/DBF4 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: CDC7/DBF4 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 CDC7/DBF4 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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