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Journal
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Biochemistry
/
NEMO IKK绑定域的生产、结晶和结构确定
JoVE Journal
Biochemistry
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JoVE Journal
Biochemistry
Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
Please note that all translations are automatically generated.
Click here for the English version.
NEMO IKK绑定域的生产、结晶和结构确定
DOI:
10.3791/60339-v
•
13:02 min
•
December 28, 2019
•
Adam H. Barczewski
,
Michael J. Ragusa
,
Dale F. Mierke
,
Maria Pellegrini
1
Department of Chemistry
,
Dartmouth College
Chapters
00:04
Title
01:12
Large Scale Expression of Histidine-6 (His
6
)-Tagged Nuclear Factor-kappa-B Essential Modulator Crystallography Construct (NEMO-EEAA)
02:33
His
6
-Tagged NEMO-EEAA Purification
04:42
His
6
Tag Cleavage and Purification
07:09
Sparse Matrix Screening
08:23
Seed Stock Generation
09:00
Crystal Looping
09:43
X-Ray Data Processing and Structure Solution
10:38
Results: Representative Production, Crystallization, and Structure Determination of the IKK-Binding Domain of NEMO
12:28
Conclusion
Summary
Automatic Translation
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Automatic Translation
我们描述了通过X射线晶体学测定NEMOIKK结合域的结构确定方案。这些方法包括蛋白质表达、纯化和表征,以及成功优化晶体和确定无结合形式蛋白质的结构的策略。
Tags
NEMO
IKK-binding Domain
Protein Production
Crystallization
Structure Determination
Coiled-coil Adapters
Protein Refolding
Protein Concentration
Aggregation
Solubility
French Press
Cell Lysis
Urea
Protein Purification
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