Histone Ubiquitin Ligase Activity Assay Service

Q: Q: Can both mono- and poly-ubiquitination be analyzed in this assay?

A: Yes. The assay design and detection method are optimized based on the ubiquitination pattern of interest.

Q: Q: Do you support assays involving E1 and E2 enzymes?

A: Yes. Complete ubiquitination cascades incorporating E1, E2, and E3 enzymes can be established as required.

Q: Q: Can endogenous ubiquitin ligase complexes be evaluated?

A: Enriched endogenous complexes can be analyzed if sufficient activity and specificity are achieved.

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

CD BioSciences offers specialized histone ubiquitin ligase activity assay services to characterize E3 ubiquitin ligase-mediated ubiquitination events on histone substrates. This service is essential for understanding the role of histone ubiquitination in chromatin remodeling, DNA damage response, and transcriptional regulation, providing critical insights for epigenetic research and therapeutic development.

Introduction to Histone Ubiquitin Ligase Activity Assay

Histone ubiquitination represents a reversible and tightly regulated epigenetic signal within the histone code. Histone ubiquitin ligases (E3 enzymes) work in concert with E1 activating enzymes and E2 conjugating enzymes to ensure site-specific ubiquitin transfer. This modification plays a pivotal role in transcription regulation, DNA repair signaling, and crosstalk with other histone marks such as methylation and acetylation.

The role of E3 ubiquitin ligases in regulating histone ubiquitination.

Fig.1 E3 ubiquitin ligases and histone ubiquitination events. (Oss-Ronen L, et al., 2022)

Activity assays for histone ubiquitin ligases are indispensable tools for elucidating enzyme function, regulatory pathways, and therapeutic relevance. By quantitatively measuring ubiquitin transfer to histone substrates, these assays support both fundamental epigenetic research and targeted inhibitor development.

Our Services

CD BioSciences offers tailored histone ubiquitin ligase activity assays using optimized substrates, reaction systems, and detection strategies. Both recombinant enzymes and enriched endogenous complexes can be analyzed to meet diverse research objectives.

Service Workflow

We offer a comprehensive and collaborative process designed to ensure accurate and reproducible results from initial consultation through final delivery. This integrated approach consists of five key phases:

Protocol Consultation and Design

Detailed understanding of client's research objectives and sample characteristics to develop personalized experimental plans, including detection method selection, experimental condition design, and expected result prediction.

Pre-experiment and Condition Optimization

Conduct small-scale pre-experiments to optimize reaction conditions, ensuring smooth main experiments and reliable results.

Formal Experiment Execution

Perform formal experiments according to optimized conditions, establishing strict control systems to ensure accuracy and reproducibility.

Data Acquisition and Analysis

Use professional software for data collection and analysis, providing detailed experimental reports and raw data.

Results Analysis & Interpretation

Provide professional interpretation of experimental results and suggestions for subsequent research directions.

Sample Requirements

E3 Ligase Samples

Purity: >80% (verified by SDS-PAGE)
Concentration: ≥0.1 mg/mL
Buffer: Provide detailed composition, avoid high concentrations of reducing agents like DTT
Storage conditions: Shipped in aliquots at -80°C

Histone Substrates

Types: Histones, nucleosomes, histone octamers
Concentration ≥1 mg/mL
Modification status: Clearly defined modification types and sites
Purity: >90%

Technical Specialization

Histone substrate expertise: Provide various histone variants and modified forms as substrates.
Complex analysis capability: Analyze different types of E3 ligase complexes (CRL, RING, HECT).
Dynamic monitoring function: Real-time tracking of ubiquitination processes for complete kinetic data.
Mechanistic research depth: Support enzyme mechanism studies, distinguishing processive vs. distributive catalytic mechanisms.

Comprehensive Application Support

Drug screening platform: Suitable for high-throughput screening of E3 ligase inhibitors
Disease model research: Support functional analysis of tumor-associated mutant E3 ligases
Integrated cross-research: Combinable with other histone modification analysis services

Quality Assurance

We are committed to providing high-quality services:

Strict experimental control setup
Repeat experiments to ensure result reliability
Detailed data recording and preservation
Timely progress communication and problem resolution

Frequently Asked Questions (FAQs)

Q: Can both mono- and poly-ubiquitination be analyzed in this assay?

Q: Do you support assays involving E1 and E2 enzymes?

Q: Can endogenous ubiquitin ligase complexes be evaluated?

Q: Is this service suitable for inhibitor screening?

CD BioSciences enables precise profiling of histone ubiquitin ligase activity to advance epigenetic research and drug discovery. Our integrated platform combines optimized assay systems with expert data interpretation to deliver actionable insights. Contact us to discuss your project goals and receive a customized experimental strategy tailored to your research needs.

Reference

1. Oss-Ronen L, Sarusi T, Cohen I. Histone mono-ubiquitination in transcriptional regulation and its mark on life: emerging roles in tissue development and disease[J]. Cells, 2022, 11(15): 2404.