Herbert B R, Woods J L
Wool Research Organisation of New Zealand, Christchurch.
Electrophoresis. 1994 Jul;15(7):972-6. doi: 10.1002/elps.11501501142.
Conventional carrier ampholyte-isoelectric focusing is unsuitable for separating wool and hair proteins. The inability to form stable narrow pH gradients at acidic pH values does not allow good resolution of the many acidic wool and hair proteins. To evaluate the usefulness of immobilised pH gradient-isoelectric focusing (IPG-IEF) for wool proteins, S-amidomethyl wool proteins were analysed using IPG-IEF. Over twenty bands were visible on a pH 4-7 IPG-IEF gel. IPG-IEF was combined with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) to give a two dimensional separation, and over 50 protein spots were observed. IPG-IEF appears to be ideally suited to the separation of wool proteins. The resolution of two-dimensional electrophoresis using IPG-IEF was greater than previously obtained using acid or alkaline PAGE in the first dimension. Good reproducibility of spot position was observed.
传统的载体两性电解质等电聚焦法不适用于分离羊毛和毛发蛋白质。在酸性pH值下无法形成稳定的窄pH梯度，这使得许多酸性羊毛和毛发蛋白质无法得到良好的分离效果。为了评估固定化pH梯度等电聚焦（IPG-IEF）对羊毛蛋白质的实用性，使用IPG-IEF分析了S-氨甲基化羊毛蛋白质。在pH 4-7的IPG-IEF凝胶上可见二十多条条带。IPG-IEF与十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）相结合进行二维分离，观察到了50多个蛋白质斑点。IPG-IEF似乎非常适合分离羊毛蛋白质。使用IPG-IEF进行二维电泳的分辨率高于之前在第一维使用酸性或碱性PAGE所获得的分辨率。观察到斑点位置具有良好的重现性。
