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진핵 세포에서 단백질 복합체의 탠덤 선호도 정화
JoVE Journal
Biyokimya
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JoVE Journal Biyokimya
Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells

진핵 세포에서 단백질 복합체의 탠덤 선호도 정화

DOI:

11:30 min

January 26, 2017

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Bölümler

  • 00:05Başlık
  • 00:52Collecting the Cells
  • 01:39Lysing the Cells
  • 02:32Equilibrating the STREP Beads
  • 03:56STREP Affinity Purification (AP)
  • 06:01Equilibrating the FLAG Beads
  • 07:04FLAG Immunoprecipitation (IP)
  • 08:39Results: A Cyclin-CDK Complex is Successfully Purified by Tandem Affinity Purification (TAP)
  • 10:26Conclusion

Özet

Otomatik Çeviri

We describe here a novel, robust, and efficient tandem affinity purification (TAP) method for the expression, isolation, and characterization of protein complexes from eukaryotic cells. This protocol could be utilized for the biochemical characterization of discrete complexes as well as the identification of novel interactors and post-translational modifications that regulate their function.

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