Abstract
Many biochemical and biophysical related questions require the isolation of functional synaptic vesicles. Isolated synaptic vesicles can be used for transporter kinetics studies, synaptic vesicle content analysis and immuno-labeling of specific synaptic vesicle proteins, etc. Here I describe a fast and reliable isolation procedure to allow researchers to isolate a large amount, as well as physiologically functional synaptic vesicles, by following the subsequent order of cryogrinding, gradient ultracentrifugation, and size exclusion liquid chromatography. This process enriches over 90% of the synaptic vesicle population, with low contamination of Golgi or endoplasmic reticulum vesicles.
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Acknowledgments
Huinan Li, Ph.D. was supported by National Institute on Aging P30AG062422-01 Grant at the time of preparing for this manuscript. This summary is part of his graduate work under the mentorship of Mark L. Harlow, Ph.D. in Department of Biology, Texas A&M University (College Station).
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Li, H. (2022). Rapid Isolation of Functional Synaptic Vesicles from Tissues Through Cryogrinding, Ultracentrifugation, and Size Exclusion Chromatography. In: Dahlmanns, J., Dahlmanns, M. (eds) Synaptic Vesicles. Methods in Molecular Biology, vol 2417. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1916-2_10
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DOI: https://doi.org/10.1007/978-1-0716-1916-2_10
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