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Генетические и биохимические подходы к<em> В Vivo</em> и<em> In Vitro</em> Оценка белка олигомеризации: The Рианодин Рецептор Case Study
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
DOI:

12:43 min

July 27, 2016

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Chapitres

  • 00:05Titre
  • 01:10Y2H: Yeast Transformation
  • 03:22Y2H: Colony-lift Filter Paper β-Gal Assay
  • 04:29Protein Expression in a Mammalian Cell Line
  • 06:56Co-immunoprecipitation
  • 09:02Chemical Cross-linking
  • 09:46Results: The Ryanodine Receptor Forms Homo-oligomers Within Cells
  • 11:40Conclusion

Summary

Traduction automatique

Oligomerization of the ryanodine receptor, a homo-tetrameric ion channel mediating Ca2+ release from intracellular stores, is critical for skeletal and cardiac muscle contraction. Here, we present complementary in vivo and in vitro methods to detect protein self-association and determine homo-oligomer stoichiometry.

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