Separation of proteins by sodium dodecylsulfate capillary electrophoresis in hydroxypropylcellulose sieving matrix with laser-induced fluorescence detection
Separation of proteins by sodium dodecylsulfate capillary electrophoresis in hydroxypropylcellulose sieving matrix with laser-induced fluorescence detection
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DOI：
10.1016/s0021-9673(00)00660-9
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发表时间：
2000-10-13
影响因子：
4.1
通讯作者：
Dovichi, NJ
中科院分区：
文献类型：
--
作者：
Hu, S;Zhang, ZR;Dovichi, NJ
建立了以羟丙基纤维素为筛分基质的十二烷基硫酸钠毛细管电泳分离蛋白质的方法。采用荧光染料3-（2-呋喃基）喹啉-2-甲醛作为柱前试剂标记蛋白质，利用激光诱导荧光提高检测灵敏度。用分子质量在14 000 ~ 97 000范围内的5种标准蛋白对该方法进行了测试，并得到了这些蛋白的分子质量与其峰值迁移时间之间的校准曲线。该方法也适用于HT29人结肠腺癌细胞提取物的蛋白质分离，通常在20分钟的分离时间内可分离近30种蛋白质成分。当使用三种运行缓冲系统时，观察到细胞提取物蛋白的分离模式相似，表明缓冲系统的组成对基于HPC筛分的分离没有太大影响。(C) 2000 Elsevier Science BN。版权所有。
Sodium dodecyl sulfate capillary electrophoresis by using hydroxypropylcellulose as the sieving matrix was developed for separation of proteins. 3-(2-furoyl)quinoline-2-carboxaldehyde, a fluorogenic dye, was used as the pre-column reagent to label proteins, which allows the use of laser-induced fluorescence to improve the detection sensitivity. Five standard proteins within the molecular mass range of 14 000-97 000 were used to test this method and a calibration curve was obtained between the molecular mass of these proteins and their peak migration times. This method was also applied to the separation of proteins from HT29 human colon adenocarcinoma cell extracts, and, typically, nearly 30 protein components could be resolved in a 20-min separation. Similar separation patterns were observed for the cell extract proteins when three running buffer systems were employed, indicating that buffer composition did not have much influence on the separation based on HPC sieving. (C) 2000 Elsevier Science BN. All rights reserved.