Co-Immunoprecipitation (Co-IP) Service

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Co-Immunoprecipitation (Co-IP) Service

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CD BioSciences provides the co-immunoprecipitation (CO-IP) service which is based on the specific interaction between antibodies and antigens, it indirectly captures interacting proteins through the bait protein-specific antibodies and recognizes physiologically relevant protein-protein interactions.

Protein-protein interactions are the main accomplisher of basic plant cell functions and are involved in almost all life activities. Therefore, the study of protein-protein interactions and the establishment of relational network maps are currently hot topics in plant protein research.

Schematic diagram of the CO-IP principle. Figure 1. Schematic diagram of the CO-IP principle. (Lin, J. S., & Lai, E. M., 2017)

Service Content

Experimental content

  • Experimental design based on specific projects, including buffers, microbeads, antibodies, etc.
  • Antibody generation and immobilization.
  • Total protein extraction and reaction with specific antibodies.
  • Capture of specific antibody-protein complexes with Protein A/G.
  • Wash to remove uncaptured proteins and obtain pure protein-antibody complexes.
  • Detection of IP results by Western Blot.
  • Data analysis and report consultation.

Advantages and disadvantages of CO-IP technology

Advantages Disadvantages
  • Prey proteins are pulled down in the natural state, which reduces the artificial interactions
  • Entire protein complexes can be pulled down, which can be used for the characterization of proteins and functional pathways.
  • Highly compatible with various sample preparation and protein identification methods
  • It may not detect low affinity and instantaneous protein-protein interactions.
  • Two proteins may bind indirectly with a third party acting as a bridge in between.
  • Predict what the prey protein is before the experiment to select the antibody.

Optimization of Co-IP experimental conditions

  • Optimize protein extraction and solubilization conditions, and screen appropriate buffers and washing solutions to improve protein solubilization and purification efficiency.
  • Optimize incubation time and temperature to improve antibody binding to target proteins.
  • Increase the washing step to remove non-specifically bound proteins.
  • Selection of blocking agents, such as bovine serum albumin (BSA) or fish protein (Gelatin), to reduce nonspecific binding.

Materials to be Provided by Clients

Plant cell > 1×107

Plant tissue > 2g/group.

Plant seeds > 3g/group.

Plant roots or fruits > 4g/group.

Target protein information (Protein A, Protein B).

Primary antibody for IP (Protein A) and its instructions.

Primary antibody for Western Blot (Protein B) and its instructions.

Delivery Reports

  • Immunoprecipitation electronic pictures and analysis results.
  • Identification of interacting proteins (Western Blot results).
  • Complete experimental procedures, instrumentation, software search parameters, etc.

CD BioSciences is a renowned biological company specializing in plant protein research and providing immunoprecipitation services. With our deep expertise in plant biology and molecular technology, we offer customized solutions for clients seeking to study protein-protein interactions in plants. If you are interested in our services, please contact us and we will help you enhance your understanding of plant biology at the molecular level.

Reference

  1. Lin, J. S., & Lai, E. M. (2017). Protein-Protein Interactions: Co-Immunoprecipitation. Methods in molecular biology (Clifton, N.J.). 1615, 211-219.

For research use only, not for clinical use.