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Journal Article

Small-scale isolation of synaptic vesicles from mammalian brain.

MPS-Authors
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Ahmed,  S.
Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society;

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Holt,  M.
Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society;

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Riedel,  D.
Facility for Electron Microscopy, MPI for biophysical chemistry, Max Planck Society;

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Jahn,  R.
Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

1739781.pdf
(Publisher version), 898KB

Supplementary Material (public)

1739781_Suppl_1.pdf
(Supplementary material), 2MB

1739781_Suppl_2.pdf
(Supplementary material), 2MB

1739781_Suppl_3.pdf
(Supplementary material), 407KB

Citation

Ahmed, S., Holt, M., Riedel, D., & Jahn, R. (2013). Small-scale isolation of synaptic vesicles from mammalian brain. Nature Protocols, 8(5), 998-1009. doi:10.1038/nprot.2013.053.


Cite as: http://hdl.handle.net/11858/00-001M-0000-000E-F986-A
Abstract
Synaptic vesicles (SVs) are essential organelles that participate in the release of neurotransmitters from a neuron. Biochemical analysis of purified SVs was instrumental in the identification of proteins involved in exocytotic membrane fusion and neurotransmitter uptake. Although numerous protocols have been published detailing the isolation of SVs from the brain, those that give the highest-purity vesicles often have low yields. Here we describe a protocol for the small-scale isolation of SVs from mouse and rat brain. The procedure relies on standard fractionation techniques, including differential centrifugation, rate-zonal centrifugation and size-exclusion chromatography, but it has been optimized for minimal vesicle loss while maintaining a high degree of purity. The protocol can be completed in less than 1 d and allows the recovery of ∼150 μg of vesicle protein from a single mouse brain, thus allowing vesicle isolation from transgenic mice.