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NEMOのIKK結合ドメインの生産・結晶化・構造決定
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
DOI:

13:02 min

December 28, 2019

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Chapters

  • 00:04Title
  • 01:12Large Scale Expression of Histidine-6 (His6)-Tagged Nuclear Factor-kappa-B Essential Modulator Crystallography Construct (NEMO-EEAA)
  • 02:33His6-Tagged NEMO-EEAA Purification
  • 04:42His6 Tag Cleavage and Purification
  • 07:09Sparse Matrix Screening
  • 08:23Seed Stock Generation
  • 09:00Crystal Looping
  • 09:43X-Ray Data Processing and Structure Solution
  • 10:38Results: Representative Production, Crystallization, and Structure Determination of the IKK-Binding Domain of NEMO
  • 12:28Conclusion

Summary

Automatic Translation

X線結晶学によるNEMOのIKK結合ドメインの構造決定のためのプロトコルについて説明する。この方法には、タンパク質発現、精製および特性評価、ならびに結晶最適化を成功させるための戦略および非結合形態のタンパク質の構造決定が含まれる。

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