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首页> 外文期刊>Brain research. Brain research protocols >Protocol for quantitative and qualitative analyses of the in vitro aggregation of synthetic beta-amyloid. A method applicable to the identification of substances that may have therapeutic efficacy for Alzheimer's disease.
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Protocol for quantitative and qualitative analyses of the in vitro aggregation of synthetic beta-amyloid. A method applicable to the identification of substances that may have therapeutic efficacy for Alzheimer's disease.

机译：合成β-淀粉样蛋白的体外聚集体定量和定性分析的协议。一种适用于鉴定对阿尔茨海默氏病可能具有治疗功效的物质的方法。

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Biochemical studies of beta-amyloid (Abeta) aggregation have been hampered by the lack of a simple method to examine simultaneously the extent of aggregation and the structural nature of the aggregates. Consequently, often only the extent of aggregation is analyzed and reported. This means that there is often no knowledge of whether the aggregates consist of fibrils, a structure crucial for their neurotoxicity. Here we describe the development of a protocol for quantitatively and qualitatively evaluating the in vitro aggregation of synthetic Abeta. Specifically, a fluorescein derivatized Abeta(1-42) peptide (FITC-Abeta(1-42)) was used as the aggregation material. We found that the fluorescent Abeta peptide aggregated as efficiently as the native ones and the extent of their aggregation could be determined accurately by fluorescence spectrophotometry. Significantly, our approach is also qualitative because it allows a direct structural examination of Abeta fibrils by fluorescence microscopy. In addition, this protocol can also be applied to the identification of substance(s) capable of inhibiting Abeta aggregation and further the assessment of the nature of such inhibition.
机译：β-淀粉样蛋白（Abeta）聚集的生化研究由于缺乏同时检查聚集程度和聚集体结构性质的简单方法而受到阻碍。因此，通常仅分析和报告聚集的程度。这意味着通常不知道聚集体是否由原纤维组成，原纤维是对其神经毒性至关重要的结构。在这里，我们描述了一种用于定量和定性评估合成Abeta体外聚集的协议的开发。具体而言，将荧光素衍生的Abeta（1-42）肽（FITC-Abeta（1-42））用作聚集材料。我们发现，荧光Abeta肽的聚集效率与天然肽一样，并且其聚集程度可以通过荧光分光光度法准确确定。重要的是，我们的方法也是定性的，因为它允许通过荧光显微镜直接对Abeta纤维进行结构检查。另外，该方案也可用于鉴定能够抑制Abeta聚集的物质，并进一步评估这种抑制的性质。

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