CO-IP Immunoprecipitation Protein Interaction Analysis
Immunoprecipitation (IP) is a technique used to isolate a specific protein from a solution using an antibody that can bind to the protein. It is one of the most widely used methods for antigen purification and detection. By adding an insoluble form of antibody-binding protein (such as protein A or Protein G conjugated to agarose beads or, more recently, magnetic beads), the antibody-protein complex is removed from the solution. Immunoprecipitation uses antigen-specific antibodies to separate target antigens from a complex antigen solution and usespolyacrylamide gel SDS-PAGE methodto analyze the number and size of antigen-reactive antibodies present in complex protein mixtures. IP allows researchers to measure the molecular weight and quantity of a given protein in a protein mixture, identify the activation state of proteins and post-translational modifications, and capture protein binding partners. Immunoprecipitation assays can detect interactions between target proteins and other proteins or nucleic acids, with Co-immunoprecipitation (Co-IP) commonly used for detecting protein interactions.
Co-IP is a powerful technique widely used to discover and detect protein-protein interactions. The principle of Co-IP is to infer interacting proteins by pulling down the target protein using antigen-antibody interactions. Specifically, the antibody against the target protein is immobilized on an affinity resin and then exposed to a designed protein mixture or cell extract. Proteins or protein complexes that interact strongly with the target protein will be co-precipitated. The identity and quantity of these interacting proteins can be identified through subsequent mass spectrometry-based interaction protein analysis.
The general procedure for Bio-Tech standard Co-IP protein interaction analysis is as follows:
• Prepare protein mixture, cell lysate, or tissue extract
• Incubate the protein mixture, cell lysate, or tissue extract with antibody-bound affinity beads
• Remove unbound proteins through multiple washing steps
• Detect interacting proteins through SDS-PAGE, western blotting, or mass spectrometry (MS) analysis
Compared to other protein interaction methods, Co-IP has the following advantages:
• The target protein is pulled down in its native state, reducing the influence of artificial factors;
• Can pull down entire protein complexes, which is useful for characterizing proteins and their functional pathways;
• Highly compatible with multiple sample preparation and protein identification methods;
• Simple and straightforward to operate.
Bio-Tech Biotechnology provides high-quality Co-IP protein interaction analysis services to customers. We offer optimized solutions for sample protein extraction, expression, and purification to ensure high-quality bait and prey proteins. Additionally, we offer customized methods for characterizing potential interacting proteins.
To save your precious time and budget, Bio-Tech offers one-stop Co-IP protein interaction analysis services, covering every step of your project, including:
Experimental design - gene synthesis - protein expression and purification - antibody cultivation and immobilization - preparation of cell lysate
Co-IP for protein interaction analysis
Interaction protein identification - data analysis - bilingual report issuance - free consultation
