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Journal
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Immunology and Infection
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分子互补亲和纯化 (BiCAP) 对多蛋白信号络合物的解剖
JoVE Journal
Immunology and Infection
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JoVE Journal
Immunology and Infection
Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)
Please note that all translations are automatically generated.
Click here for the English version.
分子互补亲和纯化 (BiCAP) 对多蛋白信号络合物的解剖
DOI:
10.3791/57109-v
•
06:45 min
•
June 15, 2018
•
Jordan F. Hastings
,
Jeremy Z.R. Han
,
Robert F. Shearer
2
,
Sean P. Kennedy
3
,
Mary Iconomou
4
,
Darren N. Saunders
,
David R. Croucher
6,7
1
The Kinghorn Cancer Centre
,
Garvan Institute of Medical Research
,
2
Ubiquitin Signaling Group, Protein Signaling Program, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences
,
University of Copenhagen
,
3
RCSI Molecular Medicine
,
Royal College of Surgeons in Ireland
,
4
Department of Epigenetics
,
Max Planck Institute of Immunobiology and Epigenetics
,
5
School of Medical Sciences
,
University of New South Wales
,
6
St Vincent’s Hospital Clinical School
,
University of New South Wales
,
7
School of Medicine and Medical Science
,
University College Dublin
Chapters
00:04
Title
00:45
Plasmid Cloning
02:54
Cell Culture and Transfection
03:41
Harvesting Lysates
04:31
Affinity Purification and Preparation of BiCAP Eluant for Western Blotting
05:22
Results: BiCAP Method is a Powerful Technique to Specifically Purify and Characterize Interacting Proteins
06:06
Conclusion
Summary
Automatic Translation
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Automatic Translation
本手稿描述了分子互补亲和纯化 (BiCAP) 的协议。这种新方法促进了任何两种相互作用的蛋白质的特定隔离和下游蛋白质组特征, 同时排除了不复杂的个体蛋白质以及与竞争的结合伙伴形成的复合体。
Tags
Bimolecular Complementation Affinity Purification
BiCAP
Protein Complexes
Signaling Pathways
Cell Behavior
Receptor Tyrosine Kinase Dimerization
Breast Cancer
GFP Nanobody
Plasmid Purification
HEK 293T Cell Transfection
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