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In vivo and in vitro Studies of Adaptor-clathrin Interaction
JoVE Journal
Biologia
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JoVE Journal Biologia
In vivo and in vitro Studies of Adaptor-clathrin Interaction
DOI:

17:14 min

January 26, 2011

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Capitoli

  • 01:34Introduction
  • 02:10Generation of GFP-tagged Sla1 and Sla1 Clatherin-binding Mutant Yeast Strains
  • 03:22Fluorescence Microscopy
  • 05:09Fluorescence Image Analysis
  • 06:41Preparation of Yeast Cytosolic Extract
  • 09:54GST-fusion Protein Affinity Assay
  • 11:53Preparation of Total Cell Extract
  • 13:36Co-immunoprecipitation Assay
  • 15:16Representative Results
  • 16:39Conclusion

Summary

Traduzione automatica

Clathrin-mediated endocytosis depends on adaptor proteins that coordinate cargo selection and clathrin coat assembly. Here we describe procedures to study adaptor-clathrin physical interaction and live cell imaging approaches using as a model the yeast endocytic adaptor protein Sla1p.

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