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的蛋白质相互作用的合作伙伴在哺乳动物细胞中使用SILAC-免疫定量蛋白质组学鉴定
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
DOI:

12:53 min

July 06, 2014

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Chapitres

  • 00:05Titre
  • 01:49Harvesting of SILAC-labeled Cell Lysates
  • 03:39Binding Lysate to Anti-GFP Beads
  • 04:45Preparation of Samples for MS Analysis
  • 06:11Data Analysis I: Understanding the Results and Removal of Low-confidence Identifications
  • 07:36Data Analysis II: Selecting High Confidence Interactions for Further Study
  • 08:58Data Analysis III: Merging Replica Datasets
  • 10:59Results: Identification of Translation Initiation Factor – Binding Proteins
  • 12:22Conclusion

Summary

Traduction automatique

SILAC免疫实验指用于发现新蛋白的有力手段:蛋白质相互作用。通过使蛋白质丰度的控制和测试样品,真相互作用的精确相对定量可以很容易地从实验的污染物区分开来,并且低亲和力相互作用,通过使用较少的严格的缓冲条件下保存。

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