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一个协议,用于基于蛋白质 - 蛋白质相互作用的鉴定<sup> 15</sup> N代谢标记,免疫沉淀法,的定量质谱法和亲和调制
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
DOI:

14:44 min

September 24, 2012

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Chapters

  • 00:05Title
  • 02:30Cell Lysis, Crosslinking and Sample Preparation
  • 05:22Immunoprecipitation
  • 07:06Sample Preparation for nano-LC-MS/MS
  • 10:50Results: HSP70B with its Nucleotide Exchange Factor CGE1
  • 14:07Conclusion

Summary

Automatic Translation

我们提出了一个快速(的定量免疫共沉淀结合击倒)的方法,前面介绍的区分真假蛋白质 - 蛋白质相互作用的变化。我们的做法是根据<sup> 15N</sup>代谢标记,ATP,免疫沉淀,并定量质谱的存在/不存在的蛋白质 - 蛋白质相互作用的亲和力的调制。

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