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Fluorescent Leakage Assay to Investigate Membrane Destabilization by Cell-Penetrating Peptide
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Biochemistry
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JoVE Journal Biochemistry
Fluorescent Leakage Assay to Investigate Membrane Destabilization by Cell-Penetrating Peptide

Fluorescent Leakage Assay to Investigate Membrane Destabilization by Cell-Penetrating Peptide

DOI:
10.3791/62028-v

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07:33 min

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December 19, 2020

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, , , ,
1PHYMEDEXP,INSERM U1046 – CNRS UMR 9214 – University of Montpellier

Chapters

  • 00:05Introduction
  • 00:53Large Unilamellar Vesicle (LUV) Preparation
  • 02:57LUV Purification
  • 03:51LUV Size and Homogeneity Characterization
  • 04:21Fluorescence Leakage Assay
  • 05:20Results: Representative Fluorescence Leakage Analysis
  • 06:34Conclusion

Summary

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The fluorescence leakage assay is a simple method that enables the investigation of peptide/membrane interactions in order to understand their involvement in several biological processes and especially the ability of cell-penetrating peptides to disturb phospholipids bilayers during a direct cellular translocation process.

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Keywords: Fluorescence Leakage AssayMembrane DestabilizationCell-penetrating PeptidesLiposomesLipid CompositionLarge Unilamellar Vesicles (LUVs)PhosphatidylcholineSphingomyelinCholesterolLipid HydrationFreeze-thawExtrusionSize Exclusion Chromatography

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