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采用碳水化合物凝胶电泳分析细胞壁多糖。

Polysaccharides Analysis of Cell Wall Using Carbohydrate Gel Electrophoresis.

机构信息

Center for Genome Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

University of Chinese Academy of Sciences, Beijing, China.

出版信息

Methods Mol Biol. 2024;2841:85-94. doi: 10.1007/978-1-0716-4059-3_7.

Abstract

The plant cell wall is rich in polysaccharides with high heterogeneity. Investigating the composition and structure of cell wall polysaccharides is crucial for understanding the functionalities of plant cell walls. Carbohydrate electrophoresis is a sensitive and rapid method to analyze polysaccharides qualitatively and quantitatively. The process includes digesting the polysaccharides with appropriate cleavage enzymes, labeling the reducing ends of the released oligosaccharides with a highly charged fluorophore, and separating the labeled oligosaccharides in a polyacrylamide gel via high-voltage electrophoresis. The generated fluorescence can be calculated as compared to that of oligosaccharide standards. Therefore, this is a convenient method for polysaccharide characterization that can be performed in most laboratories. Here, we introduce the detailed operational steps and precautions, which are helpful for researchers to quickly obtain the structural information of polysaccharides.

摘要

植物细胞壁富含具有高度异质性的多糖。研究细胞壁多糖的组成和结构对于理解植物细胞壁的功能至关重要。碳水化合物电泳是一种定性和定量分析多糖的灵敏而快速的方法。该过程包括用适当的裂解酶消化多糖,用带高电荷荧光团标记释放的寡糖的还原端,然后在聚丙烯酰胺凝胶中通过高压电泳分离标记的寡糖。产生的荧光可以与寡糖标准品进行比较。因此,这是一种方便的多糖特性分析方法,大多数实验室都可以进行。在这里,我们介绍了详细的操作步骤和注意事项,有助于研究人员快速获得多糖的结构信息。

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