Chazotte Brad
CSH Protoc. 2008 Mar 1;2008:pdb.prot4929. doi: 10.1101/pdb.prot4929.
INTRODUCTIONThe eukaryotic cell has evolved to compartmentalize its functions and transport various metabolites among cellular compartments. Therefore, in cell biology, the study of organization and structure/function relationships is of great importance. Lysosomes are membranous sacs--diverse in shape and size--containing more than 40 different acid hydrolases. The enzymes operate optimally at the acidic pH (~5) within the lysosome to break down various substances. It is thought that the highly glycosylated nature of the proteins of the Golgi membrane helps to protect them from degradation. A number of the fluorescent approaches to visualizing lysosomes make use of their acidic pH. Commonly used probes include neutral red, N-(3-[2,4-dinitrophenyl amino] propyl)-N-(3-aminopropyl)methylamine (DAMP), and acridine orange (a DNA stain). This protocol describes the labeling of lysosomes in live cells with neutral red.
引言
真核细胞已经进化出将其功能区室化并在细胞区室之间运输各种代谢物的能力。因此，在细胞生物学中，研究组织以及结构/功能关系非常重要。溶酶体是形状和大小各异的膜性囊泡，含有40多种不同的酸性水解酶。这些酶在溶酶体内的酸性pH值（约5）下发挥最佳作用，以分解各种物质。据认为，高尔基体膜蛋白的高度糖基化性质有助于保护它们不被降解。许多用于可视化溶酶体的荧光方法利用了它们的酸性pH值。常用的探针包括中性红、N-（3-[2,4-二硝基苯基氨基]丙基）-N-（3-氨基丙基）甲胺（DAMP）和吖啶橙（一种DNA染料）。本方案描述了用中性红对活细胞中的溶酶体进行标记的方法。
