Ferreira A, Eichinger D
J Immunol Methods. 1981;43(3):291-9. doi: 10.1016/0022-1759(81)90177-0.
The two-dimensional electrophoretic O'Farrell technique can be simplified in terms of effort and equipment. This simplification derives mainly from the fact that the first dimension (isoelectric focusing) is run in a vertical slab gel. Since all tracks are cast in a single template, a reproducible and uniform first dimension is obtained, allowing easy and accurate comparison among tracks after standard staining and/or radioautographic procedures. After the first dimension, the focused tracks are cut and, following equilibration, simply laid on top of the second dimensional slab gel, whose spacers are 0.1 mm thicker than those used in the first separation. Thus, basically the same equipment can be used for both dimensions. This modified technique has been experimentally validated since it has allowed the detection of structural variants of two proteins related to the mouse complement system, namely, the fourth component and the sex-limited protein.
二维电泳的奥法雷尔技术在操作和设备方面可以简化。这种简化主要源于这样一个事实，即第一维（等电聚焦）在垂直平板凝胶中进行。由于所有泳道都在单个模板中浇铸，因此可获得可重复且均匀的第一维，从而在标准染色和/或放射自显影程序后便于在泳道之间进行轻松准确的比较。在第一维之后，将聚焦的泳道切割下来，平衡后，简单地放置在第二维平板凝胶的顶部，其间隔物比第一次分离中使用的间隔物厚0.1毫米。因此，基本上可以使用相同的设备进行两个维度的操作。这种改进的技术已经通过实验验证，因为它能够检测与小鼠补体系统相关的两种蛋白质的结构变体，即第四成分和性别限制蛋白。
