Yeast Lipid Extraction and Analysis by HPTLC
Yeast Lipid Extraction and Analysis by HPTLC
复制标题
酵母脂质的提取和 HPTLC 分析
DOI：
10.21769/bioprotoc.4081
复制
发表时间：
2021
期刊：
影响因子：
0.8
通讯作者：
Zhiping Xie
中科院分区：
文献类型：
--
作者：
Dan Li;Zheng-Tan Zhang;Cheng-Wen He;Zhiping Xie
生物组织中脂质结构、性质和结合的多样性使得它们的提取和分析成为一个实验挑战。因此，即使对于最简单的单细胞真菌之一，芽生酵母(酿酒酵母)，也开发了许多提取和分析方案来分离和定量不同的分子脂物种。其中，大多数都相当复杂，很难遵循。在此，我们描述了一种产率较高的酵母总脂提取工艺，适用于后续的薄层色谱(TLC)或液质联用(LC-MS)分析。然后，我们讨论了使用最广泛的溶剂系统来分离酵母磷脂和中性脂的薄层色谱。最后，我们描述了一种简单快速的考马斯亮蓝-甲醇混合物硅胶染色方法。然后，可以使用成像软件(如ImageJ)对染色的脂类进行定量。总体而言，该协议中描述的方法是节省时间和新手友好的。
The diversity of lipid structures, properties, and combinations in biological tissues makes their extraction and analysis an experimental challenge. Accordingly, even for one of the simplest single-cellular fungi, the budding yeast (Saccharomyces cerevisiae), numerous extraction and analysis protocols have been developed to separate and quantitate the different molecular lipid species. Among them, most are quite sophisticated and tricky to follow. Herein, we describe a yeast total lipids extraction procedure with a relatively good yield, which is appropriate for subsequent thin-layer chromatography (TLC) or liquid chromatography-mass (LC-MS) analysis. We then discuss the most widely used solvent systems to separate yeast phospholipids and neutral lipids by TLC. Finally, we describe an easy and rapid method for silica gel staining by a Coomassie Brilliant Blue-methanol mixture. The stained lipid species can then be quantitated using imaging software such as ImageJ. Overall, the methods described in this protocol are time-saving and novice-friendly.