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发现蛋白质相互作用和检定蛋白质功能使用的HaloTag技术
JoVE Journal
Biology
This content is Free Access.
JoVE Journal
Biology
Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
Please note that all translations are automatically generated.
Click here for the English version.
发现蛋白质相互作用和检定蛋白质功能使用的HaloTag技术
DOI:
10.3791/51553-v
•
11:16 min
•
July 12, 2014
•
Danette L. Daniels
,
Jacqui Méndez
,
Hélène Benink
,
Andrew Niles
,
Nancy Murphy
,
Michael Ford
,
Richard Jones
,
Ravi Amunugama
,
David Allen
,
Marjeta Urh
1
Promega Corporation
,
2
MS Bioworks LLC
Chapters
00:05
Title
01:23
Protein Pulldowns: Preparation of Transiently Transfected Cells
02:50
Equilibration of the HaloLink Resin
04:01
Binding and Washing of Fusion Complexes
06:21
SDS Elution for Denaturing Gels, Western Blots, or Mass Spectrometry
07:07
Cellular Imaging of Fluorescently Labeled Fusion Proteins Using a Cytoplasmic and Nuclear Permeable Ligand
08:48
Results: Using HaloTag Technology to Determine Protein Interactions and Localization
10:47
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
的HaloTag技术是显示在小和大的蛋白质复合物从哺乳动物细胞中分离显著成功的多功能技术。在这里,我们重点介绍了这项技术的优势比现有的替代品,并展示其效用研究蛋白质功能的许多方面里面的真核细胞。
Tags
Protein Interactions
Protein Function
HaloTag Technology
Proteomics
Mass Spectrometry
Human Proteome
Protein Isolation
Multiprotein Complexes
Protein Pull-downs
Protein Localization
BRD4
HDAC1
Epigenetics
Functional Proteomics
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