河北农业大学生命科学学院,保定 071001
国家自然科学基金(31171528), 高等学校博士学科点专项科研基金(20131302110006)和河北省植物生理与病理生物学重点实验室资助项目.
College of Life Sciences,Hebei Agricultural University,Baoding 071001,China
This work was supported by The National Natural Science Foundation of China (31171528), Specialized Research Fund for the Doctoral Program of Higher Education (20131302110006) and Key Laboratory of HebeiProvince for Molecular Plant-Microbe Interaction.
蛋白质印迹技术(Western blot,WB)基于抗体的特异性了解蛋白质的表达特征. 该技术已经成为生命科学基础与应用研究的最常用技术之一. 但目前WB手工操作多、实验流程难规范、一般只能用于同一WB分析内不同样品中目标蛋白质丰度的定性或相对定量,难以在不同实验室间进行WB数据比较. 本文简要回顾了WB发展的历程,提出了升级版蛋白质印迹(WB 2.0)的概念,介绍了包括数字化、标准化、自动化、微量化、通量化以及基于内参的归一化建立数据库等为核心内容的设想. 展望了WB 2.0的应用前景,提出在WB 2.0技术大规模应用的基础上,逐步建立开放的蛋白质表达特征数据库,成为继基因组、转录组等数据库之后生命科学领域的又一支撑平台.
Western blot (WB) is widely used to investigate the expression profiling for target proteins,which depends on the specific binding with antibodies and it is a fundamental technique in basic and application field of life science. However, traditional WB technique involves many manual steps, it is difficult to set up standard operational protocols. In most of the cases, it is used only within same WB analysis for the abundance investigation of target proteins qualitatively or relative quantitatively, while problematic to carry out comparison among different laboratories. In the current paper, after a summarized historical review of WB development, the concept of advance version of WB (WB 2.0) was proposed. The key components for the design and practical steps including digitalization, standardization, automation, micro-quantification, high through-put, reference-based normalization and database establishment were presented. In perspective, the application of WB 2.0 will activate the establishment of a public accessible protein expression database, which will be another supporting platform for life science succeeding the recognized genome and transcriptome databases.
张柳,史佳楠,刘国振.蛋白质印迹技术升级版(WB 2.0)的概念与设想[J].生物化学与生物物理进展,2019,46(9):917-924
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