MtoZ Biolabs adopts a well-established yeast screening platform and has launched the Yeast Two-Hybrid screening service which can be used to screen and verify direct interactions between proteins. This service is widely applied in protein function studies, interaction network construction, and the discovery of key interaction partners, providing reliable data support for elucidating protein interaction relationships within biological systems.

What Is Yeast Two-Hybrid?

Yeast Two-Hybrid (Y2H) is a cell-based screening method used to detect direct interactions between proteins. Its principle is to fuse the proteins of interest to a transcription activation domain and a DNA-binding domain, respectively. If an interaction occurs, the reporter gene is activated and produces a detectable signal. This method enables systematic screening of interaction partners in a cellular context, making it suitable for discovering novel protein interactions and assisting in interaction network construction. It plays an important role in elucidating protein function and investigating regulatory associations.

Brückne, A. et al. International Journal of Molecular Sciences, 2009.

Figure 1. The Classical Yeast Two-Hybrid System.

Yeast Two-Hybrid Screening Service at MtoZ Biolabs

MtoZ Biolabs uses a well-established Yeast Two-Hybrid (Y2H) platform to provide clients with systematic protein interaction screening and validation services, helping identify potential interaction partners and construct key interaction networks.

• Screening for unknown protein interaction targets
• Verifying whether candidate proteins exhibit direct interactions
• Comparing interaction differences under various mutations or treatment conditions
• Assessing interaction strength to support subsequent functional studies

Our analysis offers a more comprehensive understanding of protein interaction relationships, enabling researchers to explore their biological connections in depth and advance their studies with greater efficiency.

Workflow of Yeast Two-Hybrid Screening Service

1. Vector Construction and Sample Preparation

The bait vector of the target protein is constructed, and the corresponding library or test fragments are prepared to establish the foundational conditions for interaction screening.

2. Yeast Transformation

The bait vector and library vector are transformed into yeast cells, allowing the relevant proteins to be expressed within the same system.

3. Y2H Screening

Potential interacting proteins are screened using the Yeast Two-Hybrid platform, and positive clones are identified through selection markers or reporter gene activation.

4. Positive Validation

Positive clones undergo rescreening and sequence confirmation to eliminate false positives and verify true interaction relationships.

5. Result Output

Screening and validation results are organized to generate a list of interacting proteins and a concise analysis report to support subsequent studies.

Why Choose MtoZ Biolabs?

• High-sensitivity screening: Capable of capturing potential binding signals from weak interactions or low-abundance proteins.
• High-throughput capability: Suitable for large-scale screening and can rapidly generate candidate interaction lists.
• High result reliability: Rigorous rescreening and sequence verification processes ensure the credibility of interaction results.
• Customized screening strategies: Library sources and screening conditions can be flexibly selected according to research objectives.
• One-stop service support: Covers the complete workflow from vector construction to screening, validation, and report delivery.

Applications of Yeast Two-Hybrid Screening Service

1. Signal Pathway Hierarchy Analysis

Can be used to identify interaction relationships among upstream and downstream proteins within a pathway, helping to clarify how regulatory chains are connected.

2. Mutation Impact Assessment

Analyzes the effects of amino acid substitutions or deletions on protein interaction capabilities, providing insights into the functional significance of mutations.

3. Protein Stability-Related Studies

By assessing which interaction partners enhance or weaken the stability of a target protein, this approach supports understanding of its homeostatic maintenance mechanisms.

4. Immune Regulation Interaction Studies

Can identify new interaction partners of immune receptors, signaling proteins, or regulatory factors, helping to elucidate key nodes in immune response processes.

5. Plant Regulatory Pathway Interaction Studies

Can be used to screen interaction partners of plant signaling regulatory proteins, supporting studies of growth regulation and stress response mechanisms.

Deliverables

• Comprehensive Experimental Details
• Materials, Instruments, and Methods
• Protein Interaction Analysis Results
• Visualization Charts
• Raw Data Files
• Comprehensive Analysis Report

FAQ

Q1: What types of samples are suitable?

A1: Yeast Two-Hybrid screening service is suitable for target proteins that can be stably expressed in the yeast system, including full-length proteins, functional domains, or truncated fragments. The provided protein information should be clear, well-annotated, and exhibit reliable expression characteristics. Samples should be free of contamination and degradation to ensure reliable screening results.

Q2: What is the service general workflow?

A2:

Q3: What data formats are provided?

A3: MtoZ Biolabs provides multiple standardized outputs, including:

• Raw detection files
• Protein interaction analysis results (Excel/CSV)
• Visualization charts (PNG/TIFF)
• Comprehensive analysis report (PDF, including experimental procedures, data interpretation, and interaction summary)

If special analytical requirements exist, data formats can be customized according to project specifications.

Q4: How should I prepare my samples?

A4: To ensure stable and reliable screening results, the following preparation guidelines are recommended:

• Sample purity: The submitted samples should be free of degradation and inhibitory impurities, and avoid high salt or detergents.
• Sample storage: Store long-term at -80℃ and avoid repeated freeze-thaw cycles.
• Sample transportation: Use sealed, low-temperature conditions to preserve protein structure and activity.
• Additional information: Provide sample origin, concentration, and relevant processing steps to support the development of an appropriate screening strategy.

For more information, please refer to Sample Submission Guidelines for Proteomics, Sample Submission Guidelines for Metabolomics.

Start Your Project with MtoZ Biolabs    

Contact us to discuss your experimental design or request a quote. Whether you are exploring potential protein interaction networks or conducting large-scale functional studies, MtoZ Biolabs can provide robust and professional support for your research.