Absolute Quantitation of Plant Proteins

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2018-04-02 DOI:10.1002/cppb.20064

Nagib Ahsan, Rashaun S. Wilson, Jay J. Thelen

引用次数: 3

Abstract

Among targeted proteomic techniques, AQUA-MRM is considered as one of the most reliable for accurate protein quantitation. This method displays high sensitivity, specificity, and reproducibility compared to many common biochemical techniques by coupling the use of unique, heavy-labeled peptide standards and triple-quadrupole mass spectrometry. However, there are several important steps that are required for successful development and validation of a robust AQUA-MRM assay. The following protocol outlines and details the key steps necessary for plant sample preparation as well as AQUA-MRM development and validation, specifically for absolute quantitation of plant proteins in vivo. © 2018 by John Wiley & Sons, Inc.

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植物蛋白的绝对定量

在靶向蛋白质组学技术中，AQUA-MRM被认为是最可靠的精确蛋白质定量技术之一。与许多常见的生化技术相比，该方法通过使用独特的、重标记的肽标准和三重四极杆质谱法，具有较高的灵敏度、特异性和重复性。然而，有几个重要的步骤需要成功开发和验证一个强大的AQUA-MRM分析。以下方案概述并详细介绍了植物样品制备以及AQUA-MRM开发和验证所需的关键步骤，特别是用于体内植物蛋白的绝对定量。©2018 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet 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. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.