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Bottom-up in vitro-metoder til analyse af den ultrastrukturelle organisation, membranomformning og krumningsfølsomhedsadfærd hos septiner
Journal JoVE
Biologie
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Journal JoVE Biologie
Bottom-Up In Vitro Methods to Assay the Ultrastructural Organization, Membrane Reshaping, and Curvature Sensitivity Behavior of Septins
DOI:

09:09 min

August 17, 2022

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Chapitres

  • 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

Summary

Traduction automatique

Septiner er cytoskeletale proteiner. De interagerer med lipidmembraner og kan fornemme, men også generere membrankrumning på mikronskala. Vi beskriver i denne protokol bottom-up in vitro-metoder til analyse af membrandeformationer, krumningsfølsom septinbinding og septinfilament ultrastruktur.

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