Cogliati Bruno, Maes Michaël, Pereira Isabel Veloso Alves, Willebrords Joost, Da Silva Tereza Cristina, Crespo Yanguas Sara, Vinken Mathieu
Department of Pathology, School of Veterinary Medicine and Animal Science, University of São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitária, São Paulo, SP, Brazil.
Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Jette, Belgium.
Methods Mol Biol. 2016;1437:55-70. doi: 10.1007/978-1-4939-3664-9_4.
Immunohistochemistry (IHC) is a ubiquitous used technique to identify and analyze protein expression in the context of tissue and cell morphology. In the connexin research field, IHC is applied to identify the subcellular location of connexin proteins, as this can be directly linked to their functionality. The present chapter describes a protocol for fluorescent IHC to detect connexin proteins in tissues slices and cells, with slight modifications depending on the nature of biological sample, histological processing, and/or protein expression level. Basically, fluorescent IHC is a short, simple, and cost-effective technique, which allows the visualization of proteins based on fluorescent-labeled antibody-antigen recognition.
免疫组织化学(IHC)是一种广泛应用的技术,用于在组织和细胞形态学背景下鉴定和分析蛋白质表达。在连接蛋白研究领域,免疫组织化学被用于确定连接蛋白的亚细胞定位,因为这可能与其功能直接相关。本章描述了一种用于在组织切片和细胞中检测连接蛋白的荧光免疫组织化学方法,并根据生物样品的性质、组织学处理和/或蛋白质表达水平进行了一些细微调整。基本上,荧光免疫组织化学是一种简短、简单且经济高效的技术,它基于荧光标记的抗体-抗原识别实现蛋白质的可视化。