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Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
Journal JoVE
Biologie
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Journal JoVE Biologie
Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
DOI:

08:38 min

August 14, 2011

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Chapitres

  • 00:05Titre
  • 01:10Expression and Extraction of the Proteins
  • 03:22Immobilization of ProIM and ProIMnc on the Membrane
  • 03:54Re-folding of Membrane-bound Proteins
  • 04:49Probing the ProIM by ProSOL
  • 05:33Visualizing Protein-protein Interaction by Immunoblotting
  • 07:06Representative Results
  • 08:20Conclusion

Summary

Traduction automatique

Testing protein-protein interaction is indispensable for dissection of protein functionality. Here, we introduce an in vitro protein-protein binding assay to probe a membrane-immobilized protein with a soluble protein. This assay provides a reliable method to test interaction between an insoluble protein and a protein in solution.

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