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核糖体组蛋白的体外泛化和二次基质测定
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
DOI:

11:36 min

July 25, 2019

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Chapters

  • 00:04Title
  • 01:01GSH-agarose Affinity Purification of the GST-RING1B(1-159)-BMI1 (1-109) E3 Ubiquitin Ligase Complex
  • 02:16Purification of the BAP1/DEUBAD (ASXL2) Deubiquitinase Complex
  • 04:58Purification of the Nucleosomes from HEK293T Cells
  • 07:38Nucleosome Ubiquitination Assay Using BMI1/RING1B E3 Ubiquitin Ligase Complex and In Vitro Nucleosomal H2A DUB Assay Using BAP1/DEUBAD
  • 09:05Results: Purification, Ubiquitination, and Deubiquitination
  • 10:40Conclusion

Summary

Automatic Translation

泛化是一种翻译后修饰,在细胞过程中起着重要作用,并且通过二足化进行紧密协调。两种反应的缺陷是人体疾病的基础。我们提供使用纯化成分在体外进行泛化和二次脱体反应的协议。

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