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Reconstitution of Actin-Based Motility with Commercially Available Proteins
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Reconstitution of Actin-Based Motility with Commercially Available Proteins

Reconstitution of Actin-Based Motility with Commercially Available Proteins

DOI:
10.3791/64261-v

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08:40 min

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October 28, 2022

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,
1Laboratoire de physique de l’Ecole Normale Supérieure, ENS, Université PSL, CNRS,Sorbonne Université, Université Paris Cité

Chapters

  • 00:04Introduction
  • 01:05Preparation of Protein Solutions
  • 02:04Measurement of Protein Concentrations
  • 04:15Coating of Beads
  • 05:32Preparing the Motility Mix, Slides for Observation, and Microscopy Observation
  • 06:39Results: Measuring Protein Concentrations, Analyzing Comet Formation and Bead Velocity
  • 07:53Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes how to produce actin comets on the surfaces of beads using commercially available protein ingredients. Such systems mimic the protrusive structures found in cells, and can be used to examine physiological mechanisms of force production in a simplified way.

Tags

Keywords: ActinActin-based MotilityProtein ReconstitutionProtein PurificationBiochemical AnalysisBiophysical AnalysisLangmuir EquationProtein ResuspensionProtein ConcentrationBradford Assay

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