Co-Immunoprecipitation (Co-IP)
Background
Introduction
Co-Immunoprecipitation (Co-IP) is a popular way to see how proteins team up. It works by using the specific hookup between an antibody and its target protein to fish out protein complexes from stuff like cell lysates. Basically, a lot of these protein interactions stick together under gentle conditions. Once the cells are lysed, the antibody latches onto its target, often using agarose beads. If there's another protein linked to the target inside the cell, the whole combo gets dragged down together. This method helps you see how proteins work together in the body and is a great tool for checking out protein complexes and signaling pathways.
Fig1. Principle of co-Immunoprecipitation.
Applications of Co-IP
Co-IP is really handy in biological and medical research. Here's what it's commonly used for:
- Protein Interaction Validation: It's used to confirm protein interactions you think are happening, offering clear evidence of how proteins link up in living systems.
- Discovery of New Interactors: Combined with Mass Spectrometry, Co-IP helps spot new protein partners, boosting our understanding of protein networks and how they function.
- Drug Discovery: Co-IP studies protein interactions related to drug targets, helping to find possible therapeutic candidates and figure out how drugs do their thing.
- Functional Studies: It helps uncover what protein complexes do in different cell activities, like signal transduction, gene expression, and managing the cell cycle.
Service Procedure
- Comprehensive Service Workflow:
- Deliverables:
- Detailed Experimental Report
A full report that wraps up the experimental setup, methods, and outcomes. It covers detailed data analysis too, like spotting protein interactions and confirming predicted partners. - Data Visualization
Graphs and charts illustrating the results of the Co-IP experiment, including Western Blot images or Mass Spectrometry data. This helps in visualizing the protein interactions and validating the findings. - Actionable Insights
Interpretation of the data in the context of your research goals, along with recommendations for further experiments or optimizations. We provide practical insights to help you make the most of the results.
- Detailed Experimental Report
- Service Details:
- Customized Experimental Design and Advanced Tools
We set up Co-IP experiments based on what you need, using top-notch antibodies and cutting-edge gear to make sure we catch protein interactions accurately and sensitively. Whether you're aiming to confirm protein interactions you suspect or find new ones, we'll team up with you to create a plan that fits your goals. - High-Quality Data, Detailed Reporting, and Expert Analysis
Our Co-IP services offer top-quality data and detailed reports. With techniques like Western Blot and Mass Spectrometry, we make sure you get accurate and dependable results. We include data interpretation and suggestions for further research in the report, so you can grasp the protein interactions in your study better. - Full-Process Service and Integrated Solutions
We're here to help every step of the way, from the first consultation to follow-up. Our experts guide you through the whole process to make sure your Co-IP experiment matches your needs and you get solid data and comprehensive reports. Besides Co-IP, we also offer related services like Western Blot, Mass Spectrometry, and Protein Interaction Analysis, providing all-in-one solutions for your research.
- Customized Experimental Design and Advanced Tools
Why Choose Profacgen?
Expertise and Experience
We have enough experience to know our way around protein interactions. Our team has seen it all and can design Co-IP experiments that really work.
Highly Reproducible Results
Our Co-IP services are rock-solid. We deliver results that you can rely on, every time.
Short Turn-Around Time
We recognize the importance of speed, which is why we focus on delivering results promptly without compromising quality. Your results will be ready in a timely manner.
Cost-Effective Solutions
Our pricing strategy is clear and fair. Obtaining high-quality data is affordable and won't strain your budget.
Case Study
Background
The objective of this project was to identify the interacting proteins of the target protein (XXX) in a specific cell line provided by the customer. The service involved using Co-Immunoprecipitation (Co-IP) with XXX antibody to enrich the protein complexes, followed by Mass Spectrometry (MS) to identify the specific protein types bound by XXX protein. The goal was to gain insights into the protein interactions and pathways involving XXX protein.
Results
- Western Blot Detection:
The Co-IP experiment successfully enriched proteins bound to XXX antibody. Western blot analysis confirmed the presence of specific proteins, indicating successful enrichment.
Fig2. Results of Western Blot Detection of XXX Antibody IP Enrichment.
- Mass Spectrometry Identification:
The enriched proteins were analyzed by MS, identifying multiple protein candidates interacting with XXX protein. The ion peaks and mass spectra of these proteins were recorded and analyzed.
Fig3. Mass Spectrometry Identification of Immunoprecipitation-enriched XXX Protein Ion Peaks.
- Bioinformatics Analysis:
GO Analysis: Enrichment analysis showed significant biological processes, cellular components, and molecular functions associated with the identified proteins.
Fig4. GO_BP Enrichment Analysis was Performed on Differentially Enriched Proteins by Immunoprecipitation IP of XXX Protein.
Fig5. GO_CC Enrichment Analysis was Performed on Differentially Enriched Proteins by Immunoprecipitation IP of XXX Protein.
Fig6. GO_MF Enrichment Analysis was Performed on Differentially Enriched Proteins by Immunoprecipitation IP of XXX Protein.
KEGG Pathway Analysis: Identified proteins were mapped to key metabolic and signaling pathways, providing insights into their biological roles.
Fig7. XXX Protein Co-Immunoprecipitation Enriched Differential Proteins for KEGG Enrichment Analysis.
STRING Protein Interaction Network: A protein-protein interaction network was generated, highlighting potential interactions among the identified proteins.
Fig8. XXX Protein Co-Immunoprecipitation Enriched Differential Proteins for KEGG Enrichment Analysis.
Conclusion
The project successfully identified several proteins interacting with XXX protein using Co-IP-MS. The identified proteins were further analyzed through GO, KEGG, and STRING analyses, revealing their potential roles and interactions in cellular pathways. The results provide valuable insights into the biological functions and interactions of XXX protein, achieving the project's goal.
FAQs
Resources
| Surface Plasmon Resonance (SPR) Service | Bio-Layer Interferometry (BLI) | Proximity-dependent Biotin Identification (BioID) Service | Yeast Two-Hybrid Screening | Pull Down Assay |
See more details on the principle and protocol of co-IP on our website. We also provide troubleshooting guides for the problems you may account in your experiment. See more…