Co-Immunoprecipitation

Overview

Co-immunoprecipitation (Co-IP) is a biochemical technique used to identify protein-protein interactions. An antibody specific to a bait protein is used to immunoprecipitate the target from a cell lysate; co-precipitated interacting partners are then detected by Western blot or mass spectrometry. It provides evidence of physical interaction between proteins under near-physiological conditions.

Used by

• Co-IP used to confirm direct interaction between HA-tagged SEMA7A and ITGB1 (integrin β1) in GBC cells; binding was lost when the SEMA7A RGD motif was mutated to RGE, confirming the RGD-dependent interaction mechanism PMID:24997986
• Co-IP and Co-IP mass spectrometry (PXD059301) used to identify USP10 as a TRMT10A interactor and to characterize the TRMT10A–BRCA1 interaction in the context of ATM-mediated phosphorylation PMID:28068672.

Notes

• Reciprocal Co-IP (immunoprecipitating with antibodies to both proteins) strengthens interaction claims.
• Interaction detected by Co-IP reflects association under cell-lysis conditions and does not necessarily indicate direct binding; proximity-ligation or in vitro binding assays provide complementary evidence.
• Widely used to map signaling complexes and validate protein-receptor pairings.

Sources

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