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Biology
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鉴定蛋白质相互作用的合作伙伴使用串联亲和纯化
JoVE Journal
Biology
Author Produced
This content is Free Access.
JoVE Journal
Biology
Identification of Protein Interacting Partners Using Tandem Affinity Purification
Please note that all translations are automatically generated.
Click here for the English version.
鉴定蛋白质相互作用的合作伙伴使用串联亲和纯化
DOI:
10.3791/3643-v
•
10:02 min
•
February 25, 2012
•
Dalan Bailey*
1
,
Luis Urena*
1
,
Lucy Thorne
,
Ian Goodfellow
1
Section of Virology, Department of Medicine
,
Imperial College London
Chapters
00:05
Title
01:30
Transfection
02:20
Cell Lysis
03:30
Binding to Rabbit IgG-agarose
04:53
TEV Protease Cleavage
05:48
Binding to Immobilized Streptavidin Beads
07:50
Biotin Elution of the Streptavidin Binging Peptide and Bait Protein
Summary
Automatic Translation
English (Original)
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Automatic Translation
串联亲和纯化的蛋白结合合作伙伴的标识是稳健的做法。由于概念证明,这种方法的应用以及特色的翻译起始因子eIF4E共沉淀的宿主细胞因子参与翻译起始。这种方法是很容易适应任何细胞或病毒蛋白。
Tags
Protein Interacting Partners
Tandem Affinity Purification
Identification Of Interacting Proteins
Yeast Two Hybrid System
Pull Down Assays
Immunoprecipitation
Mass Spectrometry Identification
Double-affinity Tag Mediated Purification
TAP
Eukaryotic Translation Initiation Factor EIF4E
Cellular Translation Factor EIF4E
EIF4F Complex
Cap-dependent Translation Initiation
Protein G Units
Streptavidin Binding Peptide
Tobacco Etch Virus (TEV) Protease Cleavage Sequence
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