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Preparation of Synaptic Plasma Membrane and Postsynaptic Density Proteins Using a Discontinuous Sucrose Gradient
Published on: September 3, 2014
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Preparation of synaptic plasma membrane and postsynaptic density proteins using a discontinuous sucrose gradient
Marie Kristel Bermejo1, Marija Milenkovic1, Ali Salahpour1
1Department of Pharmacology and Toxicology, University of Toronto.
Journal of Visualized Experiments : Jove
|September 17, 2014
Summary
This study details a classic method for isolating synaptic plasma membrane and post-synaptic density proteins. These neuronal fractionation techniques remain vital for quantifying synaptic proteins in neuroscience research.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Neuronal subcellular fractionation is crucial for studying protein trafficking to and from synapses.
- Established techniques from the 1960s are still widely used in neuroscience research.
Purpose of the Study:
- To detail the fractionation of synaptic plasma membrane and post-synaptic density proteins.
- To provide a method suitable for subsequent proteomic analyses.
Main Methods:
- Ultracentrifugation on a discontinuous sucrose density gradient.
- Isolation of synaptic plasma membranes.
- Subsequent isolation of detergent-insoluble post-synaptic density proteins.
Main Results:
- Successful fractionation of synaptic plasma membrane and post-synaptic density proteins.
- Protein preparations are suitable for western blotting and 2D DIGE analysis.
Conclusions:
- The described sucrose gradient fractionation method remains a fundamental technique in neuroscience.
- This method enables the quantification and analysis of key synaptic proteins.
