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

作者：Huang Chunhui; Xu Dilin; Zhang Ke; Jieyi, Shentu; Yan Sicheng; Wu Dapeng; Wang Qinwen; Cui Wei^*

来源：Jove-Journal of Visualized Experiments, 2018, (135): e57592.

DOI：10.3791/57592

• Biochemistry
• Issue 135
• Alzheimer's disease
• A beta
• oligomer
• dot blotting analysis
• transmission electronic microscopy
• curcumin

beta-amyloid (A beta) is a hydrophobic peptide with an intrinsic tendency to self-assemble into aggregates. Among various aggregates, A beta 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, A beta oligomer inhibitors might prevent neurodegeneration and have the potential to be developed as disease-modifying treatments of AD. However, different formation protocols of A beta oligomer might lead to oligomers with different characteristics. Moreover, there are not many methods to effectively screen A beta(1-42) oligomer inhibitors. An A11 antibody can react with a subset of toxic A beta(1-42) oligomer with anti-parallel beta-sheet structures. In this protocol, we describe how to prepare an A11-positive A beta(1-42) oligomer-rich sample from a synthetic A beta(1-42) peptide in vitro and to evaluate relative amounts of A11-positive A beta(1-42) oligomer in samples by a dot blotting analysis using A11 and A beta(1-42)-specific 6E10 antibodies. Using this protocol, inhibitors of A11-positive A beta(1-42) oligomer can also be screened from semi-quantitative experimental results.

• 出版日期2018-5
• 单位宁波大学