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Immunology and Infection

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Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)
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Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)

Article DOI: 10.3791/57109-v • 06:45 min • June 15th, 2018
June 15th, 2018 •
1, 1, 1,2, 1,3, 1,4, 5, 1,6,7
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

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Summary
Automatic Translation

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.

Tags

Multi-protein Signaling Complexes Bimolecular Complementation Affinity Purification (BiCAP) Cell Biology Biochemistry Protein Complexes Signaling Pathways Cell Behavior Protein-protein Interaction Tyrosine Kinase Dimerization Breast Cancer Physiological Setting Disease GFP Nanobody Fluorescent Proteins Entry Vector Destination Vector TE Buffer Recombinase ZiMex Centrifuge Proteinase K Solution Heat Shock-competent Cells
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