Hochstrasser D F, Merril C R
Section of Biochemical Genetics, National Institute of Mental Health, Bethesda, Maryland 20892.
Appl Theor Electrophor. 1988;1(1):35-40.
The crosslinker diacrylyl-piperazine produces polyacrylamide gels which display improved electrophoretic separation of proteins and better physical strength. It also produces gels with improved detection of proteins by ammoniacal silver staining by reducing the background. This reduced background provided us with an opportunity to investigate residual background staining caused by the catalytic reagents utilized in the polymerization of acrylamide gels. The commonly used catalyst system, tetramethyl-ethylenediamine and ammonium persulfate was shown to be responsible for the yellow staining background found after a prolonged development time with silver staining. An alternate catalyst system has been designed to decrease further the formation of this background staining. Dimethyl-piperazine or tetramethylethylenediamine, potassium or ammonium persulfate, and sodium thiosulfate are shown to provide for gels which have excellent mechanical and staining characteristics. These catalytic systems produce little background staining despite prolonged development time with the ammoniacal silver stain, and they reduce background staining with the dichromate silver stain.
交联剂二丙烯酰基哌嗪可制备聚丙烯酰胺凝胶,该凝胶在蛋白质电泳分离方面表现出更好的效果,且物理强度更高。它还能制备出通过氨性银染法检测蛋白质时背景更低的凝胶。这种降低的背景为我们提供了一个机会来研究丙烯酰胺凝胶聚合过程中所使用的催化试剂导致的残留背景染色。常用的催化剂体系,即四甲基乙二胺和过硫酸铵,被证明是长时间银染后出现黄色染色背景的原因。已设计出一种替代催化剂体系,以进一步减少这种背景染色的形成。二甲基哌嗪或四甲基乙二胺、过硫酸钾或过硫酸铵以及硫代硫酸钠被证明能制备出具有优异机械性能和染色特性的凝胶。尽管使用氨性银染法长时间显色,这些催化体系产生的背景染色很少,并且它们还能降低重铬酸盐银染法的背景染色。