School of Science and Technology, The Open University of Hong Kong, HKSAR, Hong Kong, China.
Int J Mol Sci. 2018 Jul 9;19(7):1998. doi: 10.3390/ijms19071998.
Although the emergence of gel-free approaches has greatly enhanced proteomic studies, two-dimensional gel electrophoresis (2-DE) remains one of the most widely used proteomic techniques for its high resolving power, relatively low cost, robustness, and high resolution. Preparation of high-quality protein samples remains the key in high-quality 2-DE for proteomic analysis. Samples with high endogenous levels of interfering molecules, such as salts, nucleic acids, lipids, and polysaccharides, would yield a low-quality 2-DE gel and hinder the analysis. Recently, a TRIzol-based protein extraction method has gained prominence and has attracted attention due to its promising performance in high-quality 2-DE. The authors evaluate the use of this approach for four valuable dried food products, namely two dried seafood products (abalone slices and whelk slices) and two traditional Chinese tonic foods (ganoderma and caterpillar fungus). The results indicate that 2-DE gels obtained through the TRIzol-based method are of high-quality and are comparable to those obtained through the trichloroacetic acid⁻acetone method in terms of spot number, spot intensity, and resolution. The TRIzol-based method is generally applicable to dried food samples and is simple and fast, which greatly streamlines the protein extraction procedure. Additionally, it enables the concurrent extraction and analysis of RNA, DNA, and protein from the same sample.
尽管无胶方法的出现极大地促进了蛋白质组学研究,但二维凝胶电泳(2-DE)仍然是最广泛使用的蛋白质组学技术之一,因为它具有高分辨率、相对低成本、稳健性和高分辨率。高质量蛋白质样品的制备仍然是蛋白质组分析高质量 2-DE 的关键。内源性干扰分子(如盐、核酸、脂质和多糖)含量高的样品会产生低质量的 2-DE 凝胶,从而阻碍分析。最近,一种基于 TRIzol 的蛋白质提取方法因其在高质量 2-DE 中的出色表现而备受关注。作者评估了该方法在四种有价值的干食品中的应用,即两种干海鲜产品(鲍鱼片和海螺片)和两种传统的中国滋补食品(灵芝和蝉花)。结果表明,基于 TRIzol 的方法获得的 2-DE 凝胶质量高,在斑点数量、斑点强度和分辨率方面可与三氯乙酸-丙酮法获得的凝胶相媲美。基于 TRIzol 的方法通常适用于干食品样品,简单快捷,极大地简化了蛋白质提取过程。此外,它还可以从同一样品中同时提取和分析 RNA、DNA 和蛋白质。