Nami Babak, Wang Zhixiang
Department of Medical Genetics and Signal Transduction Research Group, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada.
Methods Mol Biol. 2017;1652:109-116. doi: 10.1007/978-1-4939-7219-7_7.
Immunofluorescence staining is an effective method for visualizing ErbB family of receptor tyrosine kinases and to monitor their expression, phosphorylation, and localization in individual cells. However, immunofluorescence staining for membrane proteins is required preserving integrity of cell membrane intact by protecting the cells from organic solvents. Permeabilization of cells may increase nonspecific intracellular background signal as an inevitable result. This chapter describes procedures for the application of indirect immunofluorescence staining for ErbB receptors in adherent and suspended cells as well as in paraffin-embedded tissue sections.
免疫荧光染色是一种可视化受体酪氨酸激酶ErbB家族并监测其在单个细胞中的表达、磷酸化和定位的有效方法。然而，对膜蛋白进行免疫荧光染色时，需要通过保护细胞免受有机溶剂的影响来保持细胞膜的完整性。细胞通透处理不可避免地可能会增加非特异性细胞内背景信号。本章介绍了在贴壁细胞、悬浮细胞以及石蜡包埋组织切片中对ErbB受体进行间接免疫荧光染色的操作步骤。
