蛋白质阵列III：反相蛋白质阵列

Protein Arrays III: Reverse-Phase Protein Arrays.

作者信息

Yuan Yulin, Hong Xia, Lin Zuan-Tao, Wang Hongting, Heon Mikala, Wu Tianfu

机构信息

Department of Biomedical Engineering, University of Houston, 3605 Cullen Blvd, Houston, TX, 77204-5060, USA.

Department of Clinical Laboratory, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, China.

出版信息

Methods Mol Biol. 2017;1654:279-289. doi: 10.1007/978-1-4939-7231-9_21.

DOI:10.1007/978-1-4939-7231-9_21

PMID:28986799

Abstract

The reverse-phase protein array (RPPA) is to use highly specific antibodies to interrogate pan or posttranslationally modified protein targets, such as phosphorylated proteins, particularly the proteins involved in cell signaling pathways. In this protocol we will cover the preparation of cell (or tissue) lysates, sample printing, antibody validation, antibody interrogation, signal amplification steps, imaging and data analysis. In this protocol, colorimetric catalyzed signal amplification (CSA) chemistry, fluorescence and near-infrared (NIR) based detection methods will be described.

摘要

反向蛋白质阵列（RPPA）是利用高度特异性抗体来检测泛蛋白或翻译后修饰的蛋白质靶点，如磷酸化蛋白，尤其是参与细胞信号通路的蛋白质。在本方案中，我们将涵盖细胞（或组织）裂解物的制备、样本打印、抗体验证、抗体检测、信号放大步骤、成像和数据分析。在本方案中，将描述比色催化信号放大（CSA）化学、基于荧光和近红外（NIR）的检测方法。