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Sequential extraction of soluble and insoluble alpha-synuclein from Parkinsonian brains

Bandopadhyay, R; (2016) Sequential extraction of soluble and insoluble alpha-synuclein from Parkinsonian brains. Journal of visualised Experiments (JOVE) , 107 , Article e53415. 10.3791/53415. Green open access

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Abstract

Alpha-synuclein (α-syn) protein is abundantly expressed mainly within neurons, and exists in a number of different forms - monomers, tetramers, oligomers and fibrils. During disease, α-syn undergoes conformational changes to form oligomers and high molecular weight aggregates that tend to make the protein more insoluble. Abnormally aggregated α-syn is a neuropathological feature of Parkinson’s disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy (MSA). Biochemical characterisation and analysis of insoluble α-syn using buffers with increasing detergent strength and high-speed ultracentrifugation provides a powerful tool to determine the development of α-syn pathology associated with disease progression. This protocol describes the isolation of increasingly insoluble/aggregated α−syn from post-mortem human brain tissue. This methodology can be adapted with modifications to studies of normal and abnormal α-syn biology in transgenic animal models harbouring different α-syn mutations as well as in other neurodegenerative diseases that feature aberrant fibrillar deposits of proteins related to their respective pathologies.

Type: Article
Title: Sequential extraction of soluble and insoluble alpha-synuclein from Parkinsonian brains
Open access status: An open access version is available from UCL Discovery
DOI: 10.3791/53415
Publisher version: http://dx.doi.org/10.3791/53415
Language: English
Additional information: Copyright © 2016 Creative Commons Attribution 3.0 License
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Clinical and Movement Neurosciences
URI: https://discovery.ucl.ac.uk/id/eprint/1470263
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