Life Sciences Mass Spectrometry, School of Pharmaceutical Sciences, University of Geneva, CH-1211 Geneva 4, Switzerland.
Anal Biochem. 2011 Aug 15;415(2):215-7. doi: 10.1016/j.ab.2011.04.023. Epub 2011 Apr 20.
A comparative study of three detergent-free protein extraction protocols--a differential centrifugal fractionation, a delipidation protocol based on the Bligh and Dyer method, and the trifluoroethanol addition as cosolvent to an aqueous buffer--was performed on white adipose tissue. The performance of the protocols directly compatible with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was evaluated based on the total protein extraction yield and the protein recovery from different functional and cellular compartments. The most suitable method for the extraction of white adipose tissue proteins from a wide range of cellular and structural compartments was the delipidation protocol based on the Bligh and Dyer method.
对三种无去污剂蛋白提取方案(差速离心分级法、基于 Bligh 和 Dyer 法的去脂方案,以及三氟乙醇作为共溶剂添加到水性缓冲液中)在白色脂肪组织上进行了比较研究。根据总蛋白提取产率和不同功能和细胞区室的蛋白回收率,评估了与液相色谱-电喷雾电离-质谱联用(LC-ESI-MS)直接兼容的方案的性能。从广泛的细胞和结构区室中提取白色脂肪组织蛋白的最适合方法是基于 Bligh 和 Dyer 法的去脂方案。