Heukeshoven J, Dernick R
Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie, Universität Hamburg, Germany.
Electrophoresis. 1992 Sep-Oct;13(9-10):654-9. doi: 10.1002/elps.11501301137.
The preparation of homogeneous ultrathin native polyacrylamide gels, using a basic as well as an acidic buffer system is described. The basic buffer system consists of Tris-HC1/Tris-glycine, the same buffer as in sodium dodecyl sulfate (SDS)-gel electrophoresis but without SDS. The acidic system uses potassium acetate, pH 4.3, as gel buffer and beta-alanine, pH 4.6, acetic acid as electrolytes. The gels are covalently bound on glass plates. Binding of acidic gels requires a special pretreatment of glass plates. The whole procedure is simple and extraordinarily fast: 100-120 min from the start of gel preparation to the end of electrophoresis. Coomassie staining is done in 40 min and silver staining in 90 min. The native gels are excellently suited for diffusion blotting. Further attractive properties of these gels are easy handling, simple drying and dimensional stability.
本文描述了使用碱性和酸性缓冲系统制备均一超薄天然聚丙烯酰胺凝胶的方法。碱性缓冲系统由Tris-HC1/Tris-甘氨酸组成，与十二烷基硫酸钠（SDS）-凝胶电泳中的缓冲液相同，但不含SDS。酸性系统使用pH 4.3的醋酸钾作为凝胶缓冲液，pH 4.6的β-丙氨酸和醋酸作为电解质。凝胶通过共价键结合在玻璃板上。酸性凝胶的结合需要对玻璃板进行特殊预处理。整个过程简单且异常快速：从凝胶制备开始到电泳结束只需100 - 120分钟。考马斯亮蓝染色在40分钟内完成，银染色在90分钟内完成。天然凝胶非常适合扩散印迹。这些凝胶的其他吸引人的特性包括易于操作、简单干燥和尺寸稳定性。
