A11-positive &#946;-amyloid Oligomer Preparation and Assessment Using Dot Blotting Analysis

Huang Chunhui; Xu Dilin; Zhang Ke; Shentu Jieyi; Yan Sicheng; Wu Dapeng; Wang Qinwen; Cui Wei

doi:10.3791/57592

JoVE Journal
Biochemistry

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JoVE Journal Biochemistry

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

用斑点印迹分析 A11-positive β淀粉样寡聚物的制备与评价

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06:17 min

May 22, 2018

DOI: 10.3791/57592-v

Huang Chunhui^1,2, Xu Dilin¹, Zhang Ke¹, Shentu Jieyi², Yan Sicheng¹, Wu Dapeng³, Wang Qinwen¹, Cui Wei^1,2

¹Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine,Ningbo University, ²Li Dak Sum Yip Yio Chin Kenneth Li Marine Biopharmaceutical Research Center,Ningbo University, ³School of Materials Science and Chemical Engineering,Ningbo University

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本协议描述如何从合成肽体外制备 Aβ寡聚物, 并通过斑点印迹分析来评价 Aβ的相对含量。

这种方法可以帮助回答神经药理学领域的关键问题。该技术的主要优点是形成 β-淀粉样蛋白寡聚体简单、可靠且可重现。我们表明这种方法的演示至关重要，因为制备 β 寡聚体的步骤很难学习。

首先，通过向试管中加入 0.25 微升双蒸水，将 A-β 单体溶液的浓度调节至每毫升 900 毫克。然后继续用高纯度氮气蒸发单体溶液，直到体积达到约 850 微升，浓度约为 0.29 毫克/毫升。HR5P 的存在会影响 A-β 寡聚体的形成。

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