凝胶电泳蛋白质染色方法研究进展

Advances in Methods of Protein Bands Staining on SDS-PAGE

蛋白质条带染色是蛋白质凝胶电泳中一个重要的步骤。目前的方法主要有考马斯亮兰染色、银染、荧光染色、负染等。将考马斯亮兰与BBR相结合进行染色,可以减少染色/脱色的次数,能够提高考马斯亮兰在蛋白质条带上的染色效果。Gallyas Intensifying方法与银染相结合,能提高用银染已经检测到的蛋白质条带的亮度,使蛋白质点之间及其与背景之间更容易区分。将银染与考马斯亮兰结合起来进行染色可以提高同一块胶上微量蛋白质检测灵敏度。目前,对染色方法改进主要集中在如何提高蛋白质染色的灵敏度,操作方便与否,费用是否昂贵等方面。如何在保持银染方法高灵敏度的同时,又能够有效地降低染色背景,用无毒的化学物质去替换现在凝胶电泳中常用的有毒物质,克服荧光染色使蛋白质化学性质改变的缺点,在保持负染不修饰蛋白质条带化学性质优势的同时,对蛋白质条带染色结果加以改善等,可能代表了蛋白质条带染色的未来发展方向。

Protein-bands staining is an important step during the protein electrophoresis. Coomassie Brilliant Blue（CBB） staining, silver staining, fluorescent dyes staining and reverse staining were the main staining methods widely used. Combination CBB and Bismark Brown R can reduce staining/ destaining times and enhance the effect of CBB. Modified silver staining using Gallyas INTENSIFYING can increase sensitivity of protein staining to visualize faint or invisible proteins separated on silver-stained SDS- PAGE. It is easier to distinguish spots from each other and the background on the silver-stained gels. Double staining of the CBB - stained gel with silver staining can also increase the sensitivity of detection of trace proteins. The recent new methods or modification of the existed methods were mainly followed sensitivity of the stain, simpleness of the procedure and the expense. How to reduce the silver staining background, replace the toxic reagents, avoid chemical modifications in the proteins during staining and improve the reverse-staining results may be the research direction in the future.