Patterson S D, Latter G I
Cold Spring Harbor Laboratory, NY.
Biotechniques. 1993 Dec;15(6):1076-83.
The advent of storage phosphor technology has been of considerable benefit to the imaging of gel-separated radiolabeled proteins due to the rapid and quantitative nature of the data acquisition process. Previously, times over one month were required to obtain fluorographs of the same gel to yield data of sufficient dynamic range for quantitative analysis of high-resolution two-dimensional (2-D) gels. As we are in the process of building a human 2-D gel protein database, and therefore have a high throughput of 2-D gels both to image and quantitate using the Quest II software, we undertook an evaluation of a storage phosphor imager, including an evaluation of signal fade. The results of this evaluation demonstrate the feasibility of using such a system, and we describe the procedures that allow us to use this technique for quantitative analysis of many complex 2-D gel patterns. These procedures include a useful batch printing program that allows printing of many images in a non-interactive mode. Examples will be presented of how autoradiography, using storage phosphor plates and the Quest II system, have enabled us to begin building a human 2-D gel protein database including posttranslational modification information, without the previous time constraints associated with such a project.
存储磷光体技术的出现给凝胶分离的放射性标记蛋白质的成像带来了极大的便利，这得益于数据采集过程的快速性和定量性。以前，为了获得具有足够动态范围的数据以对高分辨率二维（2-D）凝胶进行定量分析，需要一个多月的时间才能得到同一块凝胶的荧光图像。由于我们正在构建一个人类二维凝胶蛋白质数据库，因此使用Quest II软件对二维凝胶进行成像和定量分析的通量很高，我们对存储磷光体成像仪进行了评估，包括对信号衰减的评估。评估结果证明了使用这种系统的可行性，我们还描述了使我们能够将该技术用于许多复杂二维凝胶图谱定量分析的程序。这些程序包括一个有用的批量打印程序，该程序允许以非交互式模式打印许多图像。将给出一些例子，说明使用存储磷光体板和Quest II系统的放射自显影技术如何使我们能够开始构建一个包含翻译后修饰信息的人类二维凝胶蛋白质数据库，而无需受此前与该项目相关的时间限制。
