GE 17513201 GLUTATHIONE SEPHAR

原核表达--GE

，容易导致目标蛋白降解，另外蛋白酶的去除也是比较烦人的问题，所以GE公司就推出了新的系列：PreScission。P系列载体是用PreScission™酶进行切割，它的消化温度只要5℃，可以最大程度减小对目的蛋白的降解作用。因为这个蛋白酶本身通过基因操作也加上了一个GST标记，所以它可以和被切下来的GST残余部分一起通过谷光甘肽凝胶柱（Glutathione Sepharose™）纯化去除。这样就可以极大程度上简化了纯化的步骤，从这里可以再次看出GE医疗产品在设计上很多时候总是考虑到下一步的纯化

GST融合蛋白的准备 Preparation of Glutathione-S-Transferase (GST) Fusion Proteins

selection Lysis buffer: PBS + 1% Triton X-100, supplemented just before use with protease inhibitors (e.g, final concentrations of 2 µg/ml aprotinin , 1 µg/ml leupeptin , and 25 µg/ml phenylmethylsulfonyl fluoride [PMSF]) Phosphate-buffered saline (PBS), ice

GST融合蛋白的准备 Preparation of Glutathione-S-Transferase (GST) Fusion Proteins

Add 5 ml of a 50:50 slurry of glutathione-Sepharose beads in lysis buffer.   Commercially available glutathione-Sepharose beads are often provided in a solution containing alcohols or other ingredients. Prior to use, these resins