Histone Kinase/Phosphatase Activity Assay Service

Q: Q: Can both kinase and phosphatase activities be measured using the same substrate?

A: In many cases, yes. Substrate compatibility depends on target residues and detection strategy. Our scientists evaluate feasibility during project design.

Q: Q: Do you support time-course or dose-response studies?

A: Yes. Kinetic analysis and dose-response profiling are available upon request.

Q: Q: Can endogenous enzymes be analyzed?

A: Enriched endogenous kinase or phosphatase preparations can be assessed if sufficient activity and specificity are present.

Q: Q: Is this service suitable for inhibitor screening?

A: Yes. The assay can be adapted for single-point testing or multi-concentration inhibitor evaluation.

Inquiry

Histone kinases and phosphatases precisely regulate phosphorylation states on serine, threonine, or tyrosine residues, making accurate activity measurement essential for functional epigenetic studies. CD BioSciences provides a sensitive and customizable in vitro platform to evaluate enzymatic phosphorylation and dephosphorylation of histone substrates under well-defined experimental conditions. We aim to delivers reliable insights into phosphorylation-driven epigenetic regulation.

Introduction to of Histone Kinase/Phosphatase Activity Assay

Histone kinases and phosphatases are key enzymes regulating histone phosphorylation. Histone kinases (e.g., Haspin) catalyze the transfer of a phosphate group from ATP to specific histone residues (e.g., serine, threonine), while histone phosphatases (e.g., PP1) remove these groups, enabling reversible, dynamic regulation. This dynamic modification is central to the "histone code," precisely controlling fundamental cellular processes like gene transcription, DNA damage repair, and mitosis by altering chromatin structure or recruiting effector complexes.

Diagram showing how histone phosphorylation functions within the DNA damage response (DDR).

Fig.1 A schematic representation of histone phosphorylation and its roles in the DNA damage response. (Gong P, et al., 2025)

Activity assays for these enzymes are crucial for elucidating their biological functions, regulatory mechanisms, and disease links. By quantifying enzymatic activity, these assays can reveal signaling pathway dysregulation and aid in targeted drug discovery. For example, phosphatase activity can be monitored in real-time using fluorogenic substrates (e.g,. DiFMUP), while immunoassays (e.g., TR-FRET) can quantify specific phosphorylation levels. These methods are essential tools for studying epigenetics-related diseases like cancer.

Our Services

CD BioSciences offers tailored activity assays to assess the catalytic function of histone kinases and phosphatases using optimized substrates and detection strategies. Assay formats are designed based on enzyme specificity, target residues, and project objectives to ensure reliable and biologically relevant results. Both recombinant enzymes and enriched endogenous preparations can be accommodated.

Radioisotopic Assays

Utilize labeled substrates (e.g., γ-³²P-ATP for kinases, ³²P-labeled histones for phosphatases) for direct, quantitative detection of phosphorylation changes.

Fluorescence-Based Methods

Employ fluorogenic substrates or antibodies for high-throughput applications with minimal background.

Luminescence Detection

Leverage luciferase-reporting systems for enhanced sensitivity in low-abundance enzyme assays.

Colorimetric Assays

Use absorbance-based detection for cost-effective screening of enzyme activities.

Detection Scope

Our assay platform supports activity analysis for:

Histone serine, threonine, or tyrosine phosphorylation.
Kinase-mediated histone phosphorylation events.
Phosphatase-mediated removal of phosphate groups.
Site-specific modifications on histones H2A, H2B, H3, and H4.
Comparative analysis of wild-type and mutant enzymes.

Custom targets and less-characterized enzymes can be evaluated upon request.

Service Workflow

Typical Applications

Functional characterization of histone kinases or phosphatases.
Comparative activity analysis of enzyme variants.
Evaluation of kinase or phosphatase inhibitors.
Mechanistic studies of phosphorylation-dependent chromatin regulation.
Support for epigenetic and signaling pathway research.

Frequently Asked Questions (FAQs)

Q: Can both kinase and phosphatase activities be measured using the same substrate?

Q: Do you support time-course or dose-response studies?

Q: Can endogenous enzymes be analyzed?

Q: Is this service suitable for inhibitor screening?

CD BioSciences enables accurate profiling of phosphorylation-related enzymatic functions in epigenetic regulation. Our platform integrates advanced methodologies for quantifying phosphorylation dynamics, supporting research in signal transduction, cell cycle regulation, and disease mechanisms. We provide end-to-end solutions tailored to diverse research needs. Contact us to discuss your research goals and receive a customized assay strategy aligned with your study objectives.

Reference

1. Gong P, Guo Z, Wang S, et al. Histone Phosphorylation in DNA Damage Response[J]. International Journal of Molecular Sciences, 2025, 26(6): 2405.