Abstract:
Subcellular fractionation is an introductory step in a variety of experimental approaches designed to study intracellular components, like membranes and organelle systems. Subcellular fractions enriched in membranes of the Golgi apparatus of mammalian cells have been isolated to address localization and activity of proteins, including enzymes, to study intracellular membrane transport mechanisms, and to reconstitute in vitro cellular processes associated with the Golgi apparatus. Here, I describe methods to purify Golgi membranes by subcellular fractionation, to assay nucleotide sulfate (PAPS) uptake into Golgi vesicles, and to measure sulfate incorporation into in vitro synthesized glycosaminoglycans.
摘要:
亚细胞分级分离是设计用于研究细胞内成分的各种实验方法的介绍性步骤。像膜和细胞器系统。已分离出富含哺乳动物细胞高尔基体膜的亚细胞部分,以解决蛋白质的定位和活性,包括酶,为了研究细胞内膜运输机制,并重建与高尔基体相关的体外细胞过程。这里,我描述了通过亚细胞分馏纯化高尔基膜的方法,为了测定高尔基体囊泡对核苷酸硫酸盐(PAPS)的摄取,并测量硫酸盐掺入到体外合成的糖胺聚糖中。