Carraro U, Rizzi C, Sandri M
CNR Unit for Muscle Biology and Physiopathology, University of Padova, Italy.
Electrophoresis. 1991 Dec;12(12):1005-10. doi: 10.1002/elps.1150121203.
A new, improved method is described, which takes advantage of the low solubility of potassium dodecyl sulfate (KDS), to concentrate sodium dodecyl sulfate-solubilized proteins from nanomolar solutions by KDS precipitation. The method was applied to proteins differing in Mr and pI. The effect of varying KCl, and the pH and/or concentration of the buffer on KDS-protein precipitation was studied. These parameters may be chosen to allow selective repartition of specific proteins in the pellet or supernatant. After precipitation, the hundred- or thousand-fold concentrated solutes are easily resuspended in small volumes of any required medium. Though initially experimented to recover muscle proteins from effluents of electroendosmotic preparative gel electrophoresis, the method proved to be of general interest as a powerful tool to recover proteins from highly diluted solutions.
本文描述了一种新的改进方法,该方法利用十二烷基硫酸钾(KDS)的低溶解度,通过KDS沉淀从纳摩尔溶液中浓缩十二烷基硫酸钠增溶的蛋白质。该方法应用于不同分子量(Mr)和等电点(pI)的蛋白质。研究了不同氯化钾浓度、缓冲液pH值和/或浓度对KDS-蛋白质沉淀的影响。可以选择这些参数,使特定蛋白质在沉淀或上清液中选择性重新分配。沉淀后,百倍或千倍浓缩的溶质很容易重新悬浮在少量任何所需的介质中。尽管最初是为了从电渗析制备凝胶电泳的流出物中回收肌肉蛋白而进行实验,但该方法被证明是一种从高度稀释溶液中回收蛋白质的有力工具,具有普遍意义。