Berkelman Tom, Bandhakavi Sricharan, Paulus Aran
Bio-Rad Laboratories, Life Science Group, Hercules, CA, USA,
Methods Mol Biol. 2015;1295:369-79. doi: 10.1007/978-1-4939-2550-6_26.
The technique of proteolytically digesting a sample and identifying its protein components by liquid chromatography followed by mass spectrometry (LC-MS) is a widely used analytical tool. Prior fractionation by isoelectric focusing (IEF) may be performed to increase the depth of proteome coverage. Here, we describe a method for in-gel IEF separation of a proteolytic digest that utilizes commercially available immobilized pH gradient (IPG) strips and a widely used IEF instrument.
通过蛋白质水解消化样品，然后利用液相色谱-质谱联用（LC-MS）鉴定其蛋白质成分的技术是一种广泛使用的分析工具。可先通过等电聚焦（IEF）进行分级分离，以增加蛋白质组覆盖深度。在此，我们描述一种利用市售固定化pH梯度（IPG）条带和广泛使用的IEF仪器对蛋白质水解消化物进行胶内IEF分离的方法。
