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Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions
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JoVE Journal Bioquímica
Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

DOI:
10.3791/54640-v

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10:44 min

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October 21, 2016

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, ,
1Departamento de Química de Biomacromoléculas, Instituto de Química,Universidad Nacional Autónoma de México, 2Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología,Universidad Nacional Autónoma de México

Capítulos

  • 00:05Título
  • 00:36SBDS-FlAsH Tag Protein Expression and Purification
  • 04:10Elongation Factor Like-1 (EFL1) Protein Expression and Purification
  • 07:07Labeling of SBDS-FlAsH with the FlAsH Fluorescent Dye 4′,5′-Bis(1,3,2 dithioarsolan-2-yl) Fluorescein and Fluorescence Anisotropy Experiments
  • 08:50Results: Evaluation of Protein-protein Interactions Using Fluorescence Anisotropy
  • 09:55Conclusion

Summary

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Tadução automática

Protein interactions are at the heart of a cell's function. Calorimetric and spectroscopic techniques are commonly used to characterize them. Here we describe fluorescence anisotropy as a tool to study the interaction between the protein mutated in the Shwachman-Diamond Syndrome (SBDS) and the Elongation factor-like 1 GTPase (EFL1).

Tags

Keywords: Fluorescence AnisotropyProtein-protein InteractionsProtein BiochemistryProtein BiophysicsSBDS-FlAsH ProteinProtein PurificationNickel Affinity ColumnSulfopropyl Cation Exchanger ColumnProtein Concentration

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