Isolating lipid droplets from multiple species
Isolating lipid droplets from multiple species
复制标题
从多个物种中分离脂滴
DOI：
10.1038/nprot.2012.142
复制
发表时间：
2013-01-01
期刊：
影响因子：
14.8
通讯作者：
Liu, Pingsheng
中科院分区：
文献类型：
--
作者：
Ding, Yunfeng;Zhang, Shuyan;Liu, Pingsheng
脂滴（LD）是一种与人类代谢综合征有关的细胞器，可用于开发生物燃料。LD的分离对于进行这种细胞器的形态学和生物化学研究至关重要。在过去的二十年中，LD已经从几种生物中分离出来，并通过显微镜，蛋白质组学和脂质组学进行了研究。然而，这些研究需要扩展到更多的模式生物，以及更多的动物组织。因此，一个标准的方法，可以很容易地应用到这些新的样本，需要最小的优化是必不可少的。在这里，我们提供了一个LD分离协议，是相对简单，适用于广泛的组织和生物体。在以往研究的基础上，该7-h方案可以产生15-100 μg蛋白质当量的高质量LD，满足目前大多数生物LD研究的要求。
The lipid droplet (LD) is a cell organelle that has been linked to human metabolic syndromes and that can be exploited for the development of biofuels. The isolation of LDs is crucial for carrying out morphological and biochemical studies of this organelle. In the past two decades, LDs have been isolated from several organisms and investigated by microscopy, proteomics and lipidomics. However, these studies need to be extended to more model organisms, as well as to more animal tissues. Thus, a standard method that can be easily applied to these new samples with the need for minimal optimization is essential. Here we provide an LD isolation protocol that is relatively simple and suitable for a wide range of tissues and organisms. On the basis of previous studies, this 7-h protocol can yield 15–100 μg of protein-equivalent high-quality LDs that satisfy the requirements for current LD research in most organisms.