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Study of Protein-protein Interactions in Autophagy Research
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Study of Protein-protein Interactions in Autophagy Research

Study of Protein-protein Interactions in Autophagy Research

DOI:
10.3791/55881-v

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14:08 min

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September 09, 2017

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, , ,
1Molecular Biology, Genetics and Bioengineering Program,Sabanci University, 2Information Center,Sabanci University, 3EFSUN, Center of Excellence for Functional Surfaces and Interfaces for Nano Diagnostics,Sabanci University

Chapters

  • 00:00Title
  • 01:25Immunofluorescence
  • 05:43Representative Results for Immunofluorescence
  • 06:16Immunoprecipitation
  • 06:38Harvesting Cells
  • 07:33Protein Isolation
  • 08:10Bradford Assay
  • 08:36Antibody Coupling of Beads
  • 09:26Incubation of Total Cell Lysate with Beads
  • 09:52Elution of Proteins from Beads
  • 10:18Preparation of Input Controls
  • 10:43Denaturation of Proteins and Western Blot
  • 13:06Representative Results for Immunoprecipitation
  • 13:29Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here are two antibody-based protein-protein interaction research techniques: immunofluorescence and immunoprecipitation. These techniques are suitable for studying physical interactions between proteins for the discovery of novel components of cellular signaling pathways and for understanding protein dynamics.

Tags

Protein-protein InteractionsAutophagyMolecular BiologyBiochemistryCell BiologyImmunoprecipitationImmunofluorescenceConfocal MicroscopyProtein LocalizationCell CultureFixationPFA

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