È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.
1Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University,Sorbonne Université, 2Institut Fresnel, CNRS UMR7249,Aix Marseille Univ, Centrale Marseille, 3Department of Bionanoscience, Kavli Institute of Nanoscience Delft,Delft University of Technology, 4Department of Chemical Engineering,Imperial College London, 5Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS),Service de microscopie électronique (IBPS-SME), 6Laboratoire Matière et Systèmes Complexes (MSC),Université Paris Cité, 7Institute of Biotechnology,Czech Academy of Sciences, BIOCEV
Capitoli
- 00:05Introduction
- 00:49Designing Wavy NOA (Norland Optical Adhesive) Replica from Wavy PDMS (Polydimethylsiloxane) Patterns
- 01:49Generation of a Supported Lipid Bilayer and Protein Incubation
- 03:22Sample Fixation, Sample Dehydration, Critical Point Drying
- 04:43Sample Mounting and Coating
- 05:54Acquire the Images Using a Field-Emission SEM (FE-SEM) Microscope
- 06:56Results: Analysis of the Self-Assembly of Septins at the Lipid Membrane
- 08:27Conclusion
Septines zijn cytoskeletale eiwitten. Ze interageren met lipidembranen en kunnen membraankromming op micronschaal waarnemen maar ook genereren. We beschrijven in dit protocol bottom-up in vitro methodologieën voor het analyseren van membraanvervormingen, krommingsgevoelige septinebinding en septinefilament ultrastructuur.
