蛋白质样品的制备和双向电泳条件是蛋白组学研究中很关键同时也是很棘手的问题。本文比较了TCA/丙酮法、酚抽提、苯酚/SDS法3种不同的蛋白提取方法,并对双向电泳的一些条件进行了优化,建立了适合萌发期大豆总蛋白的双向电泳体系,即采用17cm、p H4-7的IPG胶条,凝胶浓度为12%,蛋白质上样量为0.2mg。IEF程序为:250V 1h,500V 1h,3 000V 3h,8 000V 1h,10 000V 1h,10 000V 60 000Vh。本研究为萌发期大豆的蛋白组学研究提供了实验方法。 The preparation of protein samples and two-dimensional electrophoresis conditions are critical and also intractable problems in proteomics research. In this paper, three different extraction methods of TCA / acetone, phenol extraction and phenol / SDS were compared and some conditions of two-dimensional electrophoresis were optimized. A two-dimensional electrophoresis system suitable for total soybean protein during germination was established, IPG strips of 17 cm, p H4-7, gel concentration 12%, protein loading 0.2 mg. The IEF program is: 250V 1h, 500V 1h, 3 000V 3h, 8 000V 1h, 10 000V 1h, 10 000V 60 000Vh. This study provided experimental methods for proteomics research of germinating soybean.