Felgenhauer K
J Chromatogr. 1979 May 21;173(2):299-311. doi: 10.1016/s0021-9673(00)92298-2.
An arrangement of one- and two-dimensional polyacrylamide gradients has been applied to the analysis of multicomponent mixtures of soluble proteins: (1) two-dimensional 3-25% slabs; (2) 3-40% cylinders; and (3) 20-50% cylinders. The run in the first dimension is performed in non-restrictive agarose gel to ascertain a "pure" charge abscissa. Gradient electrophoresis is performed over prolonged periods in order to reach or closely approach the exclusion (pore) limit, which is taken as an operational parameter for the hydrodynamic size of globular molecules irrespective of their frictional ratios. For each of the three gradients one to four reference mixtures were formulated that allow size evaluations of non-denatured proteins. Human serum again proved its usefulness as a universal reference mixture for newly developed techniques. The number of stainable components is in accordance with expectation based on the broad knowledge of serum proteins. Two-dimensional agarose/gradient electrophoresis seems to fulfil the one band-one protein strategy of high-resolution techniques.
（1）二维3-25%平板；（2）3-40%柱状；（3）20-50%柱状。第一维的电泳在非限制性琼脂糖凝胶中进行，以确定一个“纯”电荷横坐标。梯度电泳长时间进行，以达到或接近排阻（孔隙）极限，该极限被视为球状分子流体力学大小的操作参数，而与它们的摩擦比无关。对于三种梯度中的每一种，都配制了一到四种参考混合物，用于对非变性蛋白质进行大小评估。人血清再次证明了其作为新开发技术通用参考混合物的有用性。可染色成分的数量与基于血清蛋白广泛知识的预期一致。二维琼脂糖/梯度电泳似乎符合高分辨率技术的一条带-一种蛋白质策略。
