Guttenberger M, Neuhoff V, Hampp R
Universität Tübingen, Institut für Botanik, Federal Republic of Germany.
Anal Biochem. 1991 Jul;196(1):99-103. doi: 10.1016/0003-2697(91)90124-c.
A method for protein determination in one- and two-dimensional electrophoresis sample buffer is presented. Accurate quantitation of protein in two-dimensional electrophoresis sample buffer (9.5 M urea, 2% Nonidet P-40, 2% carrier ampholytes, and 5% 2-mercaptoethanol) required removal of carrier ampholytes prior to the assay. This was made possible by taking advantage of the mutual solubility/insolubility of carrier ampholytes/proteins in saturated ammonium sulfate solution. In addition, improvement of protein determination in denaturing electrophoresis sample buffer containing the anionic detergent sodium dodecyl sulfate and the reducing agent 2-mercaptoethanol was achieved. The assay covers a range of sensitivity from 40 ng to 20 micrograms of protein. The procedure is applicable to large numbers of samples.
本文介绍了一种用于测定一维和二维电泳样品缓冲液中蛋白质的方法。要准确测定二维电泳样品缓冲液（9.5M尿素、2%NP-40、2%载体两性电解质和5%2-巯基乙醇）中的蛋白质，需要在测定前去除载体两性电解质。利用载体两性电解质/蛋白质在饱和硫酸铵溶液中的互溶性/不溶性可实现这一点。此外，对于含有阴离子去污剂十二烷基硫酸钠和还原剂2-巯基乙醇的变性电泳样品缓冲液中的蛋白质测定也有改进。该测定方法的灵敏度范围为40ng至20μg蛋白质。该方法适用于大量样品。
