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Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry
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Biologia
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JoVE Journal Biologia
Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry

Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry

DOI:
10.3791/53924-v

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14:47 min

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May 17, 2016

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, , , ,
1Epigenetics and Cell Fate, UMR 7216 CNRS,Centre National de la Recherche Scientifique CNRS – Université Paris Diderot, Sorbonne Paris Cité

Capítulos

  • 00:05Título
  • 00:58Isolation of Nuclei
  • 03:54Preparation of HeLa-S3 Nuclear Salt-extractable and Chromatin-bound Fractions
  • 07:05Protein Complexes Purification
  • 11:22Results: MyoD Interactors Identified by Tandem Affinity Purification Coupled to Mass Spectrometry
  • 13:37Conclusion

Summary

Tadução automática

Tadução automática

MyoD is a myogenic transcription factor with a strong capacity to induce myogenic transdifferentiation of many fully differentiated non-muscle cell lines. The epigenetic mechanisms involved in this transdifferentiation are largely unknown. Here we describe a double-affinity purification method followed by mass spectrometry to exhaustively characterize MyoD partners.

Tags

MyoDTandem Affinity PurificationMass SpectrometryInteractomeSkeletal Muscle BiologyProtein PartnersNuclear DifferentiationHeLa S3 CellsFLAG-tagged MyoDHypotonic BufferCell LysisNuclei IsolationCytoplasmic Fraction

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