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脲梯度电泳方法的技术关键
引用本文:周军贤,王希成,张艳玲,周筠梅.脲梯度电泳方法的技术关键[J].生物化学与生物物理进展,1996,23(2):172-175.
作者姓名:周军贤  王希成  张艳玲  周筠梅
作者单位:中国科学院生物物理研究所;清华大学生物科学技术系;中国科学院生物物理研究所;中国科学院生物物理研究所
摘    要:介绍在应用丙烯酸胺-脲梯度电泳技术进行蛋白质折叠、去折叠研究工作中的实验步骤和技术关键,并在文献方法的基础上作了改进。通过加入15%~0%的甘油,抵消在凝胶中由于脲浓度不同而引起的溶液粘度变化,保证在凝胶上脲浓度不同的部位对蛋白质保持同样的电泳阻力,防止前沿偏斜.采用核黄素光照催化合脲凝胶的聚合,以防止凝胶在梯度灌制完成前发生聚合.加浓缩胶和样品梳于脲梯度胶上可较好地克服边缘效应,获得好的样品迁移图谱.

关 键 词:脲梯度凝胶电泳    聚丙烯酸胺凝胶    蛋白质折叠    肌酸激酶
收稿时间:1995/5/29 0:00:00
修稿时间:1995/8/17 0:00:00

The Key Technique in Urea-gradient Electrophoresis
Zhou Junxian,Wang Xicheng,Zhao Yauling and Zhou Junmei.The Key Technique in Urea-gradient Electrophoresis[J].Progress In Biochemistry and Biophysics,1996,23(2):172-175.
Authors:Zhou Junxian  Wang Xicheng  Zhao Yauling and Zhou Junmei
Institution:Institute of Biophysics, Academia Sinica;Department of Biological Science and Biotechnology, Tsinghua University;Institute of Biophysics, Academia Sinica;Institute of Biophysics, Academia Sinica
Abstract:An improved procedure by using ureagradiont gel for protein folding and unfolding investigation is provided. Some key techniques such as urea gradient homogeneity, gel viscosity and gel edge effects have been discussed. An inverse gradient of glycerol is used to compensate for the small effects of urea on the electrophoretic mobilities of both folded and unfolded proteins. The glycerol gradient used here of 15% at 0mol/L urea to 0% at 8 mol/L urea was designed to give a constant mobility of creatine kinase at all urea concentrations. The gel was photopolymerized in the presence of riboflavin. 5% stacking gel and sample combs were also used on urea gradient gel to improve the sample pattern.
Keywords:tirea-gradient electophpresis  polyacrylamide gel  protein folding  creatine kinase
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