在多孔板中对聚丙烯酰胺中的蛋白质进行高通量蛋白质组学处理。

High-throughput proteomics processing of proteins in polyacrylamide in a multiwell format.

作者信息

Cummings Edwin D, Brown Janet M, Sarva Siva T, Waldo Robert H, Hilliard George M

机构信息

Center of Excellence in Genomics and Bioinformatics, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.

出版信息

J Proteome Res. 2007 Apr;6(4):1603-8. doi: 10.1021/pr060472y. Epub 2007 Mar 17.

DOI:10.1021/pr060472y

PMID:17367183

Abstract

Processing multiple protein samples from polyacrylamide at significant sensitivity represents a major chokepoint for raising the success rate in high-volume protein identification projects. A multiwell filterplate method for processing proteins in polyacrylamide was optimized for sensitivity using a protein standard. The results demonstrate this process to be a reliable and reproducible method over a range of gel loadings and suitable for the identification of proteins near the threshold of silver stain. This high-throughput manual method requires a minimum of specialized equipment, and can be performed disconnected from a proteomics infrastructure for the preparation of mass spectrometry-ready samples.

摘要

在高灵敏度下处理来自聚丙烯酰胺的多个蛋白质样品是提高大规模蛋白质鉴定项目成功率的一个主要瓶颈。使用蛋白质标准品对用于处理聚丙烯酰胺中蛋白质的多孔滤板方法进行了灵敏度优化。结果表明，在一系列凝胶上样量范围内，该方法是一种可靠且可重复的方法，适用于鉴定接近银染阈值的蛋白质。这种高通量手动方法所需的专用设备最少，并且可以在与蛋白质组学基础设施断开连接的情况下进行，以制备用于质谱分析的样品。