| Autor: |
Bermejo MK; Department of Pharmacology and Toxicology, University of Toronto., Milenkovic M; Department of Pharmacology and Toxicology, University of Toronto., Salahpour A; Department of Pharmacology and Toxicology, University of Toronto., Ramsey AJ; Department of Pharmacology and Toxicology, University of Toronto; a.ramsey@utoronto.ca. |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of visualized experiments : JoVE [J Vis Exp] 2014 Sep 03 (91), pp. e51896. Date of Electronic Publication: 2014 Sep 03. |
| DOI: |
10.3791/51896 |
| Abstrakt: |
Neuronal subcellular fractionation techniques allow the quantification of proteins that are trafficked to and from the synapse. As originally described in the late 1960's, proteins associated with the synaptic plasma membrane can be isolated by ultracentrifugation on a sucrose density gradient. Once synaptic membranes are isolated, the macromolecular complex known as the post-synaptic density can be subsequently isolated due to its detergent insolubility. The techniques used to isolate synaptic plasma membranes and post-synaptic density proteins remain essentially the same after 40 years, and are widely used in current neuroscience research. This article details the fractionation of proteins associated with the synaptic plasma membrane and post-synaptic density using a discontinuous sucrose gradient. Resulting protein preparations are suitable for western blotting or 2D DIGE analysis. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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