Immunofluorescence Microscopy of the Mammalian Golgi Apparatus.
Immunofluorescence Microscopy of the Mammalian Golgi Apparatus.
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哺乳动物高尔基体的免疫荧光显微镜。
DOI：
10.1007/978-1-0716-2639-9_8
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Arab M
中科院分区：
文献类型：
--
作者：
Arab M
免疫荧光是一种使用抗体和荧光团标记细胞内结构的技术。通常将细胞固定并透化，然后使用直接缀合至荧光团的第一抗体标记结构，或者更常见地，首先使用针对感兴趣的抗原的抗体，然后使用缀合至结合第一抗体的荧光团的第二抗体。荧光可以使用宽视场、共聚焦或超分辨率显微镜来可视化。在这里，我们专注于标记的高尔基体，并表明，不同的固定和透化条件可以显着影响标记的高尔基蛋白，并描述如何优化荧光检测高尔基蛋白。
Immunofluorescence is a technique that uses antibodies and fluorophores to label structures inside cells. The cells are normally fixed and permeabilized, and then structures are labelled using primary antibodies directly conjugated to fluorophores, or, more commonly, first with an antibody against an antigen of interest followed by a secondary antibody conjugated to a fluorophore that binds to the primary antibody. Fluorescence can be visualized using widefield, confocal, or super-resolution microscopy. Here we focus on labelling of the Golgi apparatus and show that different fixation and permeabilization conditions can significantly affect labelling of Golgi proteins and describe how to optimize fluorescent detection of Golgi proteins.