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A11-positive β-amyloid Oligomer Preparation and Assessment Using Dot Blotting Analysis

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title
A11-positive β-amyloid Oligomer Preparation and Assessment Using Dot Blotting Analysis
author
Chunhui, H.
author
Xu, D.
author
Zhang, K.
author
Shentu, J.
abstract
β-amyloid (Aβ) is a hydrophobic peptide with an intrinsic tendency to self-assemble into aggregates. Among various aggregates, Aβ oligomer is widely accepted as the leading neurotoxin in the progress of Alzheimer's disease (AD) and is considered to be the crucial event in the pathogenesis of AD. Therefore, Aβ oligomer inhibitors might prevent neurodegeneration and have the potential to be developed as disease-modifying treatments of AD. However, different formation protocols of Aβ oligomer might lead to oligomers with different characteristics. Moreover, there are not many methods to effectively screen Aβ1-42 oligomer inhibitors. An A11 antibody can react with a subset of toxic Aβ1-42 oligomer with anti-parallel β-sheet structures. In this protocol, we describe how to prepare an A11-positive Aβ1-42 oligomer-rich sample from a synthetic Aβ1-42 peptide in vitro and to evaluate relative amounts of A11-positive Aβ1-42 oligomer in samples by a dot blotting analysis using A11 and Aβ1-42-specific 6E10 antibodies. Using this protocol, inhibitors of A11-positive Aβ1-42 oligomer can also be screened from semi-quantitative experimental results.
subject
Aβ oligomers
date
2020-03-09T19:16:31Z (accessioned)
2020-03-09T19:16:31Z (available)
5/22/18 (issued)
identifier
Chunhui, Huang, Xu Dilin, Zhang Ke, Shentu Jieyi, Yan Sicheng, Wu Dapeng, Wang Qinwen, and Cui Wei. "A11-positive β-amyloid Oligomer Preparation and Assessment Using Dot Blotting Analysis." JoVE (Journal of Visualized Experiments) 135 (2018): e57592. (citation)
http://hdl.handle.net/10339/96054 (uri)
identifier
https://doi.org/10.3791/57592 (doi)
language
en (iso)
publisher
MyJoVE Corporation
source
JOVE
type
Article

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