Background Marine medaka is among the most popular models of fish species for ecotoxicology and environmental research and proteomic studies are useful tools for understanding the molecular responses of medaka upon exposure to different environmental stressors. The preparation of high-quality protein samples is the key to producing high-quality two-dimensional gel electrophoresis (2-DE) results for proteomic analysis. In recent years, Trizol-based protein extraction has been gaining popularity because of its promising performance in producing high-quality 2-DE as well as the convenience of the method. Methods Three Trizol-based approaches (Trizol method, Aliquot Trizol method and Trizol method with a commercial clean-up kit) were used to extract proteins from a marine medaka sample and 2-DE profiles were produced. Quality of the 2-DE profiles and effectiveness of the extraction methods were evaluated. For comparison, two common protein extraction methods (lysis buffer method and trichloroacetic acid (TCA)/acetone precipitation extraction) were also applied in parallel to Trizol-based approaches. Results Any of the three Trizol-based approaches produced a high-quality 2-DE profile of marine medaka compared with both lysis buffer method and TCA/acetone precipitation extraction. In addition, Trizol method with a commercial clean-up kit produced the best 2-DE profile in terms of background clarity, number of spots and resolution of proteins. Conclusions Trizol-based approaches offered better choices than traditional protein extraction methods for 2-DE analysis of marine medaka. The modified version of Trizol method with a commercial clean-up kit was shown to produce the best 2-DE profile.

中文翻译：

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使用基于 Trizol 的蛋白质提取方法为海洋青鳉样品制作高质量的二维凝胶电泳图谱。

背景 海洋青鳉是生态毒理学和环境研究中最受欢迎的鱼类模型之一，蛋白质组学研究是了解青鳉在暴露于不同环境压力源时的分子反应的有用工具。高质量蛋白质样品的制备是产生高质量二维凝胶电泳 (2-DE) 结果用于蛋白质组学分析的关键。近年来，基于 Trizol 的蛋白质提取因其在生产高质量 2-DE 方面的良好性能以及该方法的便利性而越来越受欢迎。方法 使用三种基于 Trizol 的方法（Trizol 方法、Aliquot Trizol 方法和带有商业净化试剂盒的 Trizol 方法）从海洋青鳉样品中提取蛋白质并生成 2-DE 谱。评估了 2-DE 配置文件的质量和提取方法的有效性。为了比较，两种常见的蛋白质提取方法（裂解缓冲液方法和三氯乙酸 (TCA)/丙酮沉淀提取）也与基于 Trizol 的方法并行应用。结果 与裂解缓冲液法和 TCA/丙酮沉淀提取法相比，三种基于 Trizol 的方法中的任何一种都产生了高质量的海洋青鳉 2-DE 剖面。此外，使用商业净化试剂盒的 Trizol 方法在背景清晰度、斑点数量和蛋白质分辨率方面产生了最佳的 2-DE 曲线。结论 基于 Trizol 的方法为海洋青鳉的 2-DE 分析提供了比传统蛋白质提取方法更好的选择。