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

7.4K Views

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06:45 min

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June 15th, 2018

DOI :

10.3791/57109-v

June 15th, 2018

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Jordan F. Hastings¹, Jeremy Z.R. Han¹, Robert F. Shearer¹^,², Sean P. Kennedy¹^,³, Mary Iconomou¹^,⁴, Darren N. Saunders⁵, David R. Croucher¹^,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

Summary

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Keywords Bimolecular Complementation Affinity Purification

Receptor Tyrosine Kinase Dimerization

Breast Cancer

GFP Nanobody

Plasmid Purification

HEK 293T Cell Transfection

Chapters in this video

0:04

Title

0:45

Plasmid Cloning

2:54

Cell Culture and Transfection

3:41

Harvesting Lysates

4:31

Affinity Purification and Preparation of BiCAP Eluant for Western Blotting

5:22

Results: BiCAP Method is a Powerful Technique to Specifically Purify and Characterize Interacting Proteins

6:06

Conclusion

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