Journal
JOURNAL OF NEUROSCIENCE METHODS
Volume 190, Issue 2, Pages 171-179Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2010.05.001
Keywords
Amyloid-beta; Atomic force microscopy; Primary neurons
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Funding
- NIH [F31-NS059283, R01-AG023593, R21-AG031878]
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Soluble A beta oligomers are recognized as playing a key role in Alzheimer's disease (AD) pathophysiology. Despite their significance, many investigators encounter difficulty generating reliable preparations for in vitro and in vivo experiments. Solutions of A beta are often unstable and soluble conformer profiles inconsistent. In this study we describe detailed methods for preparing A beta oligomers that are stable for several weeks and are enriched for low and high molecular weight oligomeric forms, including the 56-kDa form, a conformer implicated in AD-related cognitive impairment. We characterize their structural and functional properties using Western blot, dot blot, atomic force microscopy, Thioflavine T fluorescence, and primary neuronal culture toxicity assays. These synthetic preparations should prove valuable to many studying AP-mediated mechanisms underlying AD. (C) 2010 Elsevier B.V. All rights reserved.
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