利用压力循环技术（PCT）从动物微粒体中提取蛋白

The Endoplasmic Reticulum (ER) is a sub-cellular membrane network that spans the cytosol and connects the nuclear membrane to the plasma membrane. Critical cellular functions (such as protein biosynthesis, oxidation of xenobiotics, and protein transport) occur in the ER. Since ER-derived proteins account for only a small fraction of the total cellular proteome, and since the majority of ER-associated proteins are membrane proteins, which are especially difficult to purify, proteomic analysis of the ER has been a challenging endeavor. To facilitate the study of the ER proteome, an ER-like fraction of small vesicles, termed microsomes, can be isolated from cell homogenates by differential centrifugation. However, efficiently extracting proteins from this membrane-rich fraction is difficult, making a comprehensive proteomic analysis of lipid-rich microsomal samples difficult [1]. Here we describe a method for the efficient extraction of proteins from rat liver microsomes, using Pressure Cycling Technology (PCT) and the novel chemistry of the Kit. ProteoSolve 利用压力循环技术（PCT）从动物微粒体中提取蛋白 内质网(ER)是一种亚细胞膜网络结构，它位于细胞质并连接着质膜和核膜。一些重要的细胞功能比如蛋白质生物合成、异型生物质的氧化以及蛋白运输等均发生在内质网(ER)。由于起源于内质网(ER)的蛋白只占细胞蛋白总量的很小一部分，而且大多数与内质网关联的蛋白均是膜蛋白，因此这些蛋白非常难以纯化，内质网(ER)的蛋白质组学分析也因而变得非常具有挑战性。为了方便内质网(ER)蛋白的研究，一种小囊泡中类似于内质网的片段和微粒体可通过梯度离心从细胞均质中分离出来。但是，试图从这种膜含量多的片段中有效的提取蛋白是非常困难的，从而对脂质含量多的微粒体样品进行广泛的蛋白质组学分析也变得非常困难。本文描述了一种有效的从大鼠肝脏微粒体中提取蛋白的方法，该方法正是利用压力循环技术（PCT）和一种特别的ProteoSolve试剂盒。