Application of SWATH Proteomics to Mouse Biology

Q1 Agricultural and Biological Sciences Current protocols in mouse biology Pub Date : 2017-06-19 DOI:10.1002/cpmo.28

Yibo Wu, Evan G. Williams, Ruedi Aebersold

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引用次数: 10

Abstract

The quantitative measurement of the proteome has been shown to yield new insights into physiology and cell biology that cannot be determined from the genome and transcriptome because the quantitative relationship between transcriptome and proteome is complex. MS-based proteomics techniques, such as SWATH-MS, have recently advanced to the point at which they may be reliably applied by biologists who are not specialists in mass spectrometry. Here we provide standard protocols for preparation of tissue samples for input into the SWATH-MS analytical pipeline. These protocols are designed for high-throughput processing of tissues with ≥5 mg of sample available for analysis. Studies with extremely limited amounts of tissue should consider PCT-SWATH. An experienced single user should be able to process 48 samples per day for injection into the mass spectrometer, or up to 144 samples a week. The machine time necessary for running these samples with SWATH is approximately 1.5 hr per sample. Data acquisition protocols are also provided. © 2017 by John Wiley & Sons, Inc.

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SWATH蛋白质组学在小鼠生物学中的应用

由于转录组和蛋白质组之间的定量关系是复杂的，因此蛋白质组的定量测量已经被证明对生理学和细胞生物学产生了新的见解，这些见解不能从基因组和转录组中确定。基于质谱的蛋白质组学技术，如SWATH-MS，最近已经发展到可以被非质谱专家的生物学家可靠地应用的程度。在这里，我们提供了组织样品的准备标准协议，以输入到SWATH-MS分析管道。这些方案设计用于高通量处理≥5mg可用于分析的样品的组织。在组织数量极其有限的研究中，应考虑PCT-SWATH。经验丰富的单个用户应该能够每天处理48个样品以注入质谱仪，或每周多达144个样品。用SWATH运行这些样品所需的机器时间约为每个样品1.5小时。还提供了数据采集协议。©2017 by John Wiley &儿子,Inc。

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Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.