Chazotte Brad
Cold Spring Harb Protoc. 2012 Aug 1;2012(8):pdb.prot070599. doi: 10.1101/pdb.prot070599.
The Golgi may be considered a principal organizer of macromolecular traffic in the cell because many molecules, such as secreted proteins, glycoproteins, glycolipids, and plasma membrane glycoproteins, pass through the Golgi during their maturation. The fluorescent probes used to tag the Golgi make use of this function for labeling by using a fluorescent ceramide. In this protocol, two probes for labeling the Golgi are presented: a classic NBD-ceramide and a BODIPY-ceramide probe that is more resistant to photobleaching.
高尔基体可被视为细胞中大分子运输的主要组织者,因为许多分子,如分泌蛋白、糖蛋白、糖脂和质膜糖蛋白,在成熟过程中都会经过高尔基体。用于标记高尔基体的荧光探针利用这种功能,通过使用荧光神经酰胺进行标记。在本实验方案中,介绍了两种标记高尔基体的探针:一种经典的NBD-神经酰胺和一种更耐光漂白的BODIPY-神经酰胺探针。