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

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

DOI:
10.3791/2961-v

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08:38 min

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August 14, 2011

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, ,
1Department of Biochemistry and Cell Biology,State University of New York

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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.

Tags

Protein Membrane Overlay AssayInteraction Between Soluble And Insoluble ProteinsIn Vitro AssayProtein BindingBimolecular Fluorescence Complementation Assay (BiFC)Intracellular LocalizationGFP Fusion ProteinsFluorescent SignalEpifluorescence MicroscopyConfocal MicroscopyFalse Positive Results

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