@article{discovery1470263, note = {Copyright {\copyright} 2016 Creative Commons Attribution 3.0 License}, title = {Sequential extraction of soluble and insoluble alpha-synuclein from Parkinsonian brains}, journal = {Journal of visualised Experiments (JOVE)}, month = {January}, volume = {107}, year = {2016}, abstract = {Alpha-synuclein ({\ensuremath{\alpha}}-syn) protein is abundantly expressed mainly within neurons, and exists in a number of different forms - monomers, tetramers, oligomers and fibrils. During disease, {\ensuremath{\alpha}}-syn undergoes conformational changes to form oligomers and high molecular weight aggregates that tend to make the protein more insoluble. Abnormally aggregated {\ensuremath{\alpha}}-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 {\ensuremath{\alpha}}-syn using buffers with increasing detergent strength and high-speed ultracentrifugation provides a powerful tool to determine the development of {\ensuremath{\alpha}}-syn pathology associated with disease progression. This protocol describes the isolation of increasingly insoluble/aggregated {\ensuremath{\alpha}}?syn from post-mortem human brain tissue. This methodology can be adapted with modifications to studies of normal and abnormal {\ensuremath{\alpha}}-syn biology in transgenic animal models harbouring different {\ensuremath{\alpha}}-syn mutations as well as in other neurodegenerative diseases that feature aberrant fibrillar deposits of proteins related to their respective pathologies.}, url = {http://dx.doi.org/10.3791/53415}, author = {Bandopadhyay, R}, issn = {1940-087X} }