Semi-denaturing detergent agarose gel electrophoresis (SDD-AGE) was proposed by Vitaly V. Kushnirov in the Michael D. Ter-Avanesyan's laboratory as a method to compare sizes of amyloid aggregates. Currently, this method is widely used for amyloid investigation, but mostly as a qualitative approach. In this work, we assessed the possibilities and limitations of the quantitative analysis of amyloid aggregate size distribution using SDD-AGE results. For this purpose, we used aggregates of two well-characterized yeast amyloid-forming proteins, Sup35 and Rnq1, and developed a protocol to standardize image analysis and process the result. A detailed investigation of factors that may affect the results of SDD-AGE revealed that both the cell lysis method and electrophoresis conditions can substantially affect the estimation of aggregate size. Despite this, quantitative analysis of SDD-AGE results is possible when one needs to estimate and compare the size of aggregates on the same gel, or even in different experiments, if the experimental conditions are tightly controlled and additional standards are used.

中文翻译：

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用半变性洗涤剂琼脂糖凝胶电泳估计淀粉样蛋白聚集体的大小及其局限性。

半变性洗涤剂琼脂糖凝胶电泳 (SDD-AGE) 由 Vitaly V. Kushnirov 在 Michael D. Ter-Avanesyan 的实验室提出，作为比较淀粉样蛋白聚集体大小的一种方法。目前，这种方法广泛用于淀粉样蛋白调查，但主要是作为一种定性方法。在这项工作中，我们评估了使用 SDD-AGE 结果对淀粉样蛋白聚集体大小分布进行定量分析的可能性和局限性。为此，我们使用了两种已充分表征的酵母淀粉样蛋白形成蛋白 Sup35 和 Rnq1 的聚合体，并开发了一个协议来标准化图像分析和处理结果。对可能影响 SDD-AGE 结果的因素的详细调查表明，细胞裂解方法和电泳条件都会对聚集体大小的估计产生重大影响。