University of Pittsburgh, Center for Biologic Imaging, Pittsburgh, Pennsylvania.
University of Pittsburgh, Department of Surgery, Pittsburgh, Pennsylvania.
Curr Protoc. 2022 Sep;2(9):e549. doi: 10.1002/cpz1.549.
Immunohistochemistry is an essential technique for the localization and measurement of proteins in cells and tissues. This article describes methods for labeling proteins in adherent and suspension cell cultures and in tissue sections. Choices of antibodies and detection methods are discussed, and detailed troubleshooting guidelines are provided. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Immunofluorescent labeling of cells grown as adherent monolayers Alternate Protocol 1: Immunofluorescent labeling of cells in suspension Basic Protocol 2: Immunofluorescent labeling of tissue sections Alternate Protocol 2: Immunofluorescent labeling using streptavidin-biotin conjugates Alternate Protocol 3: Immunofluorescent double-labeling of tissue sections Alternate Protocol 4: Immunofluorescent double-labeling of tissue sections with two primary antibodies from the same host species.
免疫组织化学是定位和测量细胞和组织中蛋白质的重要技术。本文描述了用于贴壁和悬浮细胞培养物以及组织切片中标记蛋白质的方法。讨论了抗体和检测方法的选择,并提供了详细的故障排除指南。© 2022 年 Wiley 期刊 LLC. 基本方案 1:贴壁单层培养细胞的免疫荧光标记 可选方案 1:悬浮细胞的免疫荧光标记 基本方案 2:组织切片的免疫荧光标记 可选方案 2:使用链霉亲和素-生物素缀合物的免疫荧光标记 可选方案 3:组织切片的免疫荧光双重标记 可选方案 4:来自同一宿主物种的两种一抗的免疫荧光双重标记组织切片