Tastet Christophe, Lescuyer Pierre, Diemer Hélène, Luche Sylvie, van Dorsselaer Alain, Rabilloud Thierry
CEA-Laboratoire de Bioénergétique Cellulaire et Pathologique, Grenoble, France.
Electrophoresis. 2003 Jun;24(11):1787-94. doi: 10.1002/elps.200305400.
A new, versatile, multiphasic buffer system for high-resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins in the relative molecular weight range of 300 000-3000 Da is described. The system, based on the theory of multiphasic zone electrophoresis, allows complete stacking and destacking of proteins in the above M(r) range. The buffer system uses taurine and chloride as trailing and leading ion, respectively, and Tris, at a pH close to its pK(a), as the buffering counterion. Coupled with limited variation in the acrylamide concentration, this electrophoresis system allows to tailor the resolution in the 6-200 kDa M(r) range, with minimal difficulties in the post electrophoretic identification processes.
本文描述了一种新型的、通用的多相缓冲系统,用于在相对分子量范围为300000 - 3000 Da的蛋白质进行高分辨率十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳。该系统基于多相区带电泳理论,能使上述分子量范围内的蛋白质完全堆积和解堆积。该缓冲系统分别使用牛磺酸和氯离子作为尾随离子和前导离子,并使用pH接近其pKa的Tris作为缓冲抗衡离子。结合丙烯酰胺浓度的有限变化,这种电泳系统能够在6 - 200 kDa分子量范围内调整分辨率,且在电泳后鉴定过程中遇到的困难最小。