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Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
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Biochemie
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JoVE Journal Biochemie
Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

DOI:
10.3791/55498-v

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09:35 min

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April 01, 2017

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, , ,
1Proteomic Mass Spectrometry,Wellcome Trust Sanger Institute

Kapitel

  • 00:05Titel
  • 00:40Isolation of Native Protein Complexes by FLAG Affinity Purification
  • 03:10Blue Native PAGE
  • 04:39Preparation for Mass Spectrometry
  • 08:06Results: Characterization of Two Distinct Mta2-containing NuRD Complexes by ABC-MS
  • 08:54Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

Here we present protocols for affinity purification of protein complexes and their separation by blue native PAGE, followed by protein correlation profiling using label free quantitative mass spectrometry. This method is useful to resolve interactomes into distinct protein complexes.

Tags

Keywords: Affinity PurificationBlue Native PAGEProtein Correlation ProfilingProtein ComplexesFLAG-tagged ProteinCell LysisProtein PurificationElutionMass Spectrometry

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