Harrington M G, Zewert T E
Beckman Institute, California Institute of Technology, Pasadena 91125.
Electrophoresis. 1994 Feb;15(2):195-9. doi: 10.1002/elps.1150150133.
The properties of poly(ethylene glycol)methacrylate-acrylamide copolymer media (PEGMACs) were exploited in two ways: (i) in the first-dimensional gel of two-dimensional electrophoresis (2-DE) of hydrophobic proteins and (ii) for high speed, high-resolution electrophoresis at low temperatures. In the first application, improved resolution and yield of isoelectric focusing (IEF) separations for the hydrophobic protein zein was achieved compared to IEF in standard polyacrylamide gels. This appears promising as a candidate approach for higher resolution 2-DE mapping of uncharacterized hydrophobic proteins. In the second application, PEGMACs compatible with hydroorganic antifreeze buffer systems below cooling of the gels to low temperatures (-20 degrees C), which allowed greater current to be tolerated during electrophoresis. PEGMAC gels enabled us to perform sixfold faster electrophoretic separations and achieve threefold improved resolution of six standard proteins at the lower temperatures in a direct comparison with the normal sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system. If this approach is coupled with more precise instrumentation to control low temperatures during electrophoresis, greater separation speeds and resolution may be anticipated.
聚（甲基丙烯酸乙二醇酯 - 丙烯酰胺）共聚物介质（PEGMACs）的特性有两种应用方式：（i）用于疏水性蛋白质二维电泳（2-DE）的第一维凝胶中，以及（ii）用于低温下的高速、高分辨率电泳。在第一种应用中，与在标准聚丙烯酰胺凝胶中进行等电聚焦（IEF）相比，疏水性蛋白质玉米醇溶蛋白的IEF分离的分辨率和产率得到了提高。作为对未表征的疏水性蛋白质进行更高分辨率2-DE图谱分析的候选方法，这似乎很有前景。在第二种应用中，PEGMACs与低于凝胶冷却至低温（-20℃）的有机防冻缓冲系统兼容，这使得在电泳过程中能够承受更大的电流。与正常的十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS-PAGE）系统直接比较，PEGMAC凝胶使我们能够在较低温度下以快六倍的速度进行电泳分离，并使六种标准蛋白质的分辨率提高三倍。如果这种方法与更精确的仪器相结合以在电泳过程中控制低温，则可能预期有更高的分离速度和分辨率。
