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Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)
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1The Kinghorn Cancer Centre,Garvan Institute of Medical Research, 2Ubiquitin Signaling Group, Protein Signaling Program, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences,University of Copenhagen, 3RCSI Molecular Medicine,Royal College of Surgeons in Ireland, 4Department of Epigenetics,Max Planck Institute of Immunobiology and Epigenetics, 5School of Medical Sciences,University of New South Wales, 6St Vincent’s Hospital Clinical School,University of New South Wales, 7School of Medicine and Medical Science,University College Dublin
Chapters
- 00:04Title
- 00:45Plasmid Cloning
- 02:54Cell Culture and Transfection
- 03:41Harvesting Lysates
- 04:31Affinity Purification and Preparation of BiCAP Eluant for Western Blotting
- 05:22Results: BiCAP Method is a Powerful Technique to Specifically Purify and Characterize Interacting Proteins
- 06:06Conclusion
This manuscript describes the protocol for Bimolecular Complementation Affinity Purification (BiCAP). This novel method facilitates the specific isolation and downstream proteomic characterization of any two interacting proteins, while excluding un-complexed individual proteins as well as complexes formed with competing binding partners.
