Abramovitz A S, Randolph V, Mehra A, Christakos S
Prep Biochem. 1984 Aug;14(3):205-21. doi: 10.1080/10826068408070629.
A technique for high yield recovery of native, biologically active proteins from preparative polyacrylamide gel slices by reverse polarity elution is described. No apparatus other than the standard slab gel electrophoresis system is required. Several proteins have been recovered in biologically active form at a 90% yield, in quantities ranging from 0.4 mg to 4.2 mg. The method is effective with both small (9,000 dalton) and large (186,000 dalton) polypeptides. Both simple and complex proteins are recovered intact. For example, the copper-zinc and manganese superoxide dismutases from crude soybean extracts are active upon recovery. Similarly, the vitamin D-dependent calcium binding proteins from rat kidney and intestine are isolated by this method in homogeneous, active form.
本文描述了一种通过反相洗脱从制备型聚丙烯酰胺凝胶切片中高产率回收天然生物活性蛋白质的技术。除了标准的平板凝胶电泳系统外，无需其他仪器。已以90%的产率回收了几种生物活性形式的蛋白质，数量从0.4毫克到4.2毫克不等。该方法对小（9000道尔顿）和大（186000道尔顿）的多肽均有效。简单和复杂的蛋白质都能完整回收。例如，粗大豆提取物中的铜锌和锰超氧化物歧化酶回收后具有活性。同样，通过该方法从大鼠肾脏和肠道中分离出的维生素D依赖性钙结合蛋白呈均一的活性形式。
