c-Myc Antibodies: Unveiling New Mechanisms of Protein Interaction in Gene Expression Regulation

1. Literature Information

Research Focus: Exploration of c-Myc’s interaction with HBXIP, validation of their direct binding, identification of key interaction domains, and the functional role of the protein complex in regulating gene transcription—with c-Myc antibodies as core research tools.

Core Innovation: Discovery of a novel regulatory mechanism where HBXIP enhances c-Myc’s transcriptional activity via direct interaction (key domain: HBXIP aa56-173), revealing a positive feedback loop in oncogene expression—validated through multi-technique collaboration, with c-Myc antibodies enabling precise detection and interaction verification.

2. Research Background

c-Myc, a key proto-oncogenic transcription factor, regulates gene expression involved in cell proliferation, metabolism, and tumorigenesis. Its function is tightly controlled by protein interactions, but the underlying mechanisms remain incompletely understood. HBXIP, a regulatory protein, was found to co-localize with c-Myc in the nucleus of breast cancer cells, suggesting a potential functional interaction. c-Myc antibodies serve as indispensable tools for verifying this interaction, identifying binding domains, and exploring the complex’s role in transcriptional regulation—addressing gaps in understanding oncogene regulatory networks.

3. Research Approaches

To dissect the c-Myc-HBXIP interaction mechanism, the research team adopted a multi-technique, systematic strategy:

1. Co-localization Analysis: Using immunofluorescence (IF) with c-Myc antibodies to confirm nuclear co-localization of c-Myc and HBXIP in MCF-7 cells.
2. Interaction Validation: Employing co-immunoprecipitation (Co-IP) and GST pull-down assays to verify direct physical binding.
3. Domain Mapping: Identifying key interaction domains via truncated HBXIP fragments and mutation analysis.
4. Functional Assessment: Using electrophoretic mobility shift assays (EMSA) and luciferase reporter assays to evaluate the complex’s impact on transcriptional activity.
5. Cross-Validation: Integrating multiple techniques to ensure reliability of interaction and functional conclusions.

4. Research Outcomes

4.1 Protein Interactions in Oncogene Expression Regulation

• c-Myc’s Functional Dependence: Its transcriptional activity is regulated not only by expression levels but also by interactions with regulatory proteins (e.g., HBXIP).
• Nuclear Co-localization: IF staining with c-Myc antibodies confirms HBXIP and c-Myc co-localize in the nucleus of MCF-7 cells—providing spatial evidence for functional interaction.

4.2 Validation of Direct HBXIP-c-Myc Interaction

• Co-IP Assays: Bidirectional verification (HBXIP as bait precipitates c-Myc; c-Myc as bait precipitates HBXIP) confirms in vivo interaction.
• GST Pull-Down Assays: In vitro binding of GST-HBXIP to His-c-Myc excludes cellular co-factors, validating direct molecular interaction.

4.3 Identification of Key Interaction Domains

• HBXIP Truncation Analysis: Only full-length HBXIP and the aa56-173 fragment bind c-Myc (Co-IP/GST pull-down validation).
• Critical Motif: Mutation of the leucine chain in HBXIP abrogates binding to c-Myc—highlighting its role in structural integrity for interaction.

4.4 Transcriptional Regulation by the HBXIP-c-Myc Complex

• EMSA Assays: The complex binds DNA via c-Myc’s E-box sequence; addition of HBXIP antibody induces a super-shift, confirming HBXIP’s participation in the DNA-binding complex.
• Luciferase Reporter Assays: HBXIP dose-dependently enhances c-Myc’s transcriptional activation—establishing a positive feedback loop.

4.5 Multi-Technique Collaborative Verification

4.6 Biological Significance

• Mechanistic Insight: Reveals a new regulatory pathway for c-Myc, where HBXIP enhances its transcriptional activity—promoting oncogene expression in tumors.
• Therapeutic Potential: The HBXIP-c-Myc interaction interface (HBXIP aa56-173) serves as a target for developing novel cancer therapies.
• Methodological Paradigm: Demonstrates the power of integrated techniques for protein interaction and function research.

5. Product Empowerment by ANT BIO PTE. LTD.

ANT BIO PTE. LTD.’s STARTER brand, a leader in recombinant antibodies, provides a core tool for this research: the "S-RMab® c-Myc Recombinant Rabbit Monoclonal Antibody" (Catalog No.: S0B2165).

Key Roles of the Product:

1. Interaction Verification: Enables Co-IP and GST pull-down assays to confirm HBXIP-c-Myc binding.
2. Localization & Expression Detection: Clear nuclear staining in FFPE samples (IHC/IF) supports subcellular co-localization studies.
3. Functional Research: Facilitates EMSA and luciferase assays to evaluate transcriptional regulation.
4. Core Advantages: High specificity, minimal batch variation, and multi-platform validation (IHC, WB, IF)—ensuring reliable results for basic and translational research.

ANT BIO PTE. LTD.’s portfolio includes phospho-c-Myc (Ser62) antibodies (S0B6390) for studying post-translational regulation of c-Myc.

6. Brand Mission

ANT BIO PTE. LTD. is dedicated to empowering global life science advancement through three specialized sub-brands: ABSIN (general reagents, ELISA kits), STARTER (antibodies), and UA (recombinant proteins). Leveraging advanced development platforms—including recombinant rabbit/mouse monoclonal antibody generation, PTM Pan-Modification Antibody Platform, and One-Step ELISA—we deliver high-quality, compliant products certified by EU 98/79/EC, ISO9001, and ISO13485. Our mission is to partner with research institutions, pharmaceutical companies, and scientists worldwide, providing innovative reagents and solutions that accelerate discoveries in cancer biology, gene regulation, and precision medicine.

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8. AI Disclaimer

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ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.