Validating Protein Interactions with Co-Immunoprecipitation Using Endogenous and Tagged Protein Models

Immunoprecipitation (IP) leverages the specificity of an antibody to enrich proteins in a sample and investigate its interactions with other macromolecules. This presentation includes a general workflow for co-immunoprecipitation (co-IP) and what you should consider before setting up your experiment, including:

• Choice of model system;
• Choice of cell lysis buffers and when to include a pre-clear step;
• Primary antibody dilution and choice of beads;
• How to mitigate IgG masking by heavy and light chains;
• Using epitope tags for IP: advantages and key optimization considerations;
• Controls for IP experiments targeting post-translational modifications.

For more expert guidance on setting up your IP experiment, watch this Tech Tips video: How to Select an Antibody for Immunoprecipitation