Lill Jennie R, Nesatyy Victor J
Genentech, Inc., South San Francisco, CA, USA.
Methods Mol Biol. 2012;869:521-32. doi: 10.1007/978-1-61779-821-4_46.
Rapid evolution of state-of-the-art proteomic analyses has encompassed development of high-throughput analytical instrumentation and bioinformatic tools. However, recently, there has been a particular emphasis on increasing the throughput of sample preparation, which has become one of the rate-limiting steps in protein characterization workflows. Researchers have been investigating alternative methods to conventional convection oven incubations to try and reduce sample preparation time for protein characterization. Several protocols have appeared in the literature, which employ microwave irradiation as a tool for the preparation of biological samples for subsequent characterization by a variety of analytical techniques. In this chapter, techniques for microwave-assisted protein staining, destaining, and digestion are described. In general, the application of microwave-assisted technologies resulted in the drastic reduction of overall sample preparation time, though discrepancies in the reproducibility of several published digestion protocols still remain to be clarified.
最先进的蛋白质组学分析技术的快速发展涵盖了高通量分析仪器和生物信息学工具的开发。然而，最近特别强调提高样品制备的通量，这已成为蛋白质表征工作流程中的限速步骤之一。研究人员一直在研究替代传统对流烘箱孵育的方法，以尝试减少蛋白质表征的样品制备时间。文献中出现了几种方案，这些方案采用微波辐射作为制备生物样品的工具，以便随后通过各种分析技术进行表征。在本章中，描述了微波辅助蛋白质染色、脱色和消化的技术。一般来说，微波辅助技术的应用显著减少了整体样品制备时间，不过几个已发表的消化方案的重现性差异仍有待阐明。
