TWO-DIMENSIONAL POLYACRYLAMIDE-GEL ELECTROPHORESIS OF RIBOSOMAL-PROTEINS IN THE NANOGRAM RANGE
TWO-DIMENSIONAL POLYACRYLAMIDE-GEL ELECTROPHORESIS OF RIBOSOMAL-PROTEINS IN THE NANOGRAM RANGE
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DOI：
10.1515/bchm3.1985.366.2.901
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发表时间：
1985-01-01
期刊：
影响因子：
--
通讯作者：
KAMP, RM
中科院分区：
文献类型：
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作者：
BROCKMOLLER, J;KAMP, RM
描述了一种二维凝胶电泳系统，用于识别和检查来自不同生物体的纳克级核糖体蛋白的纯度。该程序结合了 Geyl 等人的方法。用于分离大肠杆菌核糖体蛋白，以及 Neuhoff 和 Poehling 描述的蛋白质微量电泳系统，并进行了一些修改。第一个凝胶尺寸在毛细管中进行，第二个凝胶尺寸以平板凝胶的形式进行，两者都在新设计的适合 10-20 个样品的室中运行。该电泳系统能够将运行时间从 2 天缩短至 2 小时，并通过考马斯蓝染色将灵敏度从正常 100 倍的 3-5 μg 提高。 100 mm 凝胶为 50-100 ng。所有核糖体蛋白在微凝胶上的分辨率（30×38×0.5毫米）与在微凝胶上100×的分离相似。 100次3 毫米，如 Geyl 等人所述。
A two-dimensional gel electrophoresis system to identify and check the purity of ribosomal proteins from different organisms with nanogram quantities is described. This procedure combines the method of Geyl et al. for the separation of ribosomal proteins of Escherichia coli, and the microscale electrophoresis system for proteins described by Neuhoff and Poehling, with several modifications. The first gel dimension is carried out in capillaries and the second in the form of slab gels, both are run in newly designed chambers suitable for 10-20 samples. This electrophoresis system enables a reduction of the running time from 2 days to 2 hours and an increase in sensitivity, with Coomassie blue staining, from 3-5 .mu.g for the normal 100 .times. 100 mm gels to 50-100 ng. The resolution of all ribosomal proteins on the micro-gel (30 .times. 38 .times. 0.5 mm) is similar to the separation on the mini-gel of 100 .times. 100 .times. 3 mm as described by Geyl et al.