Developing Protocols of Tricine-SDS-PAGE for Separation of Polypeptides in the Mass Range 1-30 kDa with Minigel Electrophoresis System

作者：Jiang, Shunyan; Liu, Shuqin; Zhao, Chunjiang^*; Wu, Changxin

来源：International Journal of Electrochemical Science, 2016, 11(1): 640-649.

• Low molecular weight protein
• Minigel
• Tricine-SDS-PAGE
• Western Blot

An efficient method for the separation of proteins in the mass range 1-100 kDa with minigel electrophoresis system was developed in the present study. Tricine (N-(2-Hydroxy-1,1-bis (hydroxymethyl) ethyl) glycine)-SDS-PAGE allows a resolution for proteins smaller than 30 kDa at lower acrylamide concentration than that in other electrophoretic methods. But reliable Tricine-SDS-PAGE protocols for minigel system remain to be established. In our study, Tricine-SDS-PAGEs were conducted with different composition of gels varied with concentration of acrylamide-bisacrylamide, urea, and glycerol, to develop optimized protocols of Tricine-SDS gels for minigel system. Our results indicated that gel composition containing 10% glycerol (w/v) and 4.2M urea could provide an ideal resolution for separation of small mass proteins. Then calibration curves for different types of separating gels demonstrated effective linear mobilities, and correlation coefficients of all gels were above 0.95, and the highest is the 10%T, 3%C gel, which reached 0.97. Results from the immunoblotting experiment of a 14-kDa mouse HMGA2 polypeptide also showed that the optimized protocols of minigel had excellent compatibility with Western Blot. In conclusion, the developed protocols of Tricine-SDS-PAGE for minigel provide a new choice for efficient, reproducible and convenient separation of low molecular weight proteins.

• 出版日期2016-1
• 单位中国农业大学