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Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain

Subcellular Fractionation for the Isolation of Synaptic Components from the Murine Brain

DOI:
10.3791/64574-v

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12:14 min

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September 14, 2022

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, ,
1Department of Neurology,Yale University, 2Department of Neuroscience,Yale University, 3Interdepartmental Neuroscience Program,Yale University

Chapters

  • 00:04Introduction
  • 01:37Mouse Brain Excision
  • 03:32Synaptosome Preparation
  • 05:23Hypotonic Lysis
  • 06:24Synaptic Vesicle Isolation
  • 07:35Synaptic Plasma Membrane Isolation
  • 10:02Results: Electron Microscopy of Synaptosomes and Immunoblot Analysis of Subcellular Fractions
  • 11:00Conclusion

Summary

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This protocol presents a robust, detailed method to obtain highly pure synaptosomes, synaptic vesicles, and other synaptic fractions from the mouse brain. This method enables the evaluation of synaptic processes, including the biochemical analysis of protein localization and function with compartmental resolution.

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Keywords: Subcellular FractionationSynapsesSynaptosomeBrain DisordersProtein DistributionProtein ActivityBuffer Preparation

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