通过蛋白质印迹法分析纳米材料毒性

Analysis of Nanomaterial Toxicity by Western Blot.

作者信息

Long Gao, Mo Yiqun, Zhang Qunwei, Jiang Mizu

机构信息

Department of Gastroenterology, Affiliated Children's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China.

Department of Environmental and Occupational Health Sciences, School of Public Health and Information Sciences, University of Louisville, Louisville, KY, USA.

出版信息

Methods Mol Biol. 2019;1894:161-169. doi: 10.1007/978-1-4939-8916-4_10.

DOI:10.1007/978-1-4939-8916-4_10

PMID:30547461

Abstract

Western blot is a routine biochemical technique for the immunodetection of proteins in cells and tissues exposed to nanomaterials (NMs). It is a sensitive method for protein analysis that involves the solubilization and separation of proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferring and immobilizing proteins onto a solid support, and targeted immunoprobing of a specific antigen. As a convenient and reliable research tool, the western blot plays an irreplaceable role in the era of proteomics together with mass spectrometry and protein chip. In this chapter we describe the detailed protocol for the entire process from sample preparation to quantitative measurement of target proteins.

摘要

蛋白质印迹法是一种常规生化技术，用于免疫检测暴露于纳米材料（NMs）的细胞和组织中的蛋白质。它是一种灵敏的蛋白质分析方法，包括通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）溶解和分离蛋白质，将蛋白质转移并固定到固相支持物上，以及对特定抗原进行靶向免疫检测。作为一种方便可靠的研究工具，蛋白质印迹法在蛋白质组学时代与质谱分析和蛋白质芯片一起发挥着不可替代的作用。在本章中，我们描述了从样品制备到目标蛋白质定量测量的整个过程的详细方案。