Journal
/
Neuroscience
/
Preparation of Synaptic Plasma Membrane and Postsynaptic Density Proteins Using a Discontinuous Sucrose Gradient
JoVE Journal
Neuroscience
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Neuroscience
Preparation of Synaptic Plasma Membrane and Postsynaptic Density Proteins Using a Discontinuous Sucrose Gradient

Preparation of Synaptic Plasma Membrane and Postsynaptic Density Proteins Using a Discontinuous Sucrose Gradient

DOI:
10.3791/51896-v

•

08:06 min

•

September 03, 2014

•

, , ,
1Department of Pharmacology and Toxicology,University of Toronto

Chapters

  • 00:05Title
  • 01:16Tissue Homogenization and Fractioning
  • 03:57Isolating the Synaptic Plasma Membrane (SPM) Fraction
  • 07:02Western Blot of Fractions
  • 07:32Conclusion

Summary

Automatic Translation

Automatic Translation

This article details the enrichment of proteins associated with the synaptic plasma membrane by ultracentrifugation on a discontinuous sucrose gradient. The subsequent preparation of post-synaptic density proteins is also described. Protein preparations are suitable for western blotting or 2D DIGE analysis.

Tags

Synaptic Plasma MembranePostsynaptic DensitySucrose GradientNeuronal Subcellular FractionationSynaptic Protein IsolationMembrane Protein IsolationPostsynaptic Density Protein IsolationWestern Blotting2D DIGE

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/51896?article_type=v

Studying Synaptic Vesicle Pools using Photoconversion of Styryl Dyes
check_url/51896?article_type=v

Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises
check_url/51896?article_type=v

Quantitative Analysis of Synaptic Vesicle Pool Replenishment in Cultured Cerebellar Granule Neurons using FM Dyes
check_url/51896?article_type=v

Preparation of Synaptoneurosomes from Mouse Cortex using a Discontinuous Percoll-Sucrose Density Gradient
check_url/51896?article_type=v

A General Method for Evaluating Incubation of Sucrose Craving in Rats
check_url/51896?article_type=v

Lateral Diffusion and Exocytosis of Membrane Proteins in Cultured Neurons Assessed using Fluorescence Recovery and Fluorescence-loss Photobleaching
check_url/51896?article_type=v

Post-embedding Immunogold Labeling of Synaptic Proteins in Hippocampal Slice Cultures
check_url/51896?article_type=v

Deriving the Time Course of Glutamate Clearance with a Deconvolution Analysis of Astrocytic Transporter Currents
check_url/51896?article_type=v

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy
check_url/51896?article_type=v

Simultaneous Two-photon In Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex

Read Article