Roberts Michael D, Young Kaelin C, Fox Carlton D, Vann Christopher G, Roberson Paul A, Osburn Shelby C, Moore Johnathon H, Mumford Petey W, Romero Matthew A, Beck Darren T, Haun Cody T, Badisa Veera L D, Mwashote Benjamin M, Ibeanusi Victor, Kavazis Andreas N
School of Kinesiology, Auburn University, Auburn, AL 36849, USA.
Department of Cell Biology and Physiology, Edward Via College of Osteopathic Medicine Auburn Campus, Auburn, AL 36849, USA.
J Biol Methods. 2020 Feb 24;7(1):e127. doi: 10.14440/jbm.2020.307. eCollection 2020.
Several published protocols exist for isolating contractile or myofibrillar (MF) proteins from skeletal muscle, however, achieving complete resuspension of the myofibril pellet can be technically challenging. We performed several previously published MF isolation methods with the intent of determining which method was most suitable for MF protein isolation and solubilization. Here, we provide an optimized protocol to isolate sarcoplasmic and solubilized MF protein fractions from mammalian skeletal muscle suitable for several downstream assays.
目前已有多种公开的从骨骼肌中分离收缩蛋白或肌原纤维(MF)蛋白的方案,然而,要使肌原纤维沉淀完全重悬在技术上具有挑战性。我们实施了几种先前发表的MF分离方法,目的是确定哪种方法最适合MF蛋白的分离和溶解。在此,我们提供了一种优化方案,用于从哺乳动物骨骼肌中分离肌浆蛋白和可溶解的MF蛋白组分,适用于多种下游分析。