Mason John N, Tomlinson Ian D, Rosenthal Sandra J, Blakely Randy D
Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN, USA.
Methods Mol Biol. 2005;303:35-50. doi: 10.1385/1-59259-901-X:035.
The quantum dot is a novel fluorescent platform that has the potential to become an alternative to conventional organic dyes used to label biological probes such as antibodies or ligands. Compared to typical fluorescent organic dyes, cadmium selenide/zinc sulfide core-shell nanocrystals, or quantum dots, have greater photostability, resist metabolic and chemical degradation, are nontoxic, and display broad emission and narrow excitation bands. When conjugated to generic adaptor molecules such as streptavidin, quantum dots can be used to label different biotinylated antibodies or ligands without having to customize the quantum dot surface chemistry for each antibody or ligand. In this chapter, we outline the methodology for using streptavidin quantum dots to label biotinylated antibodies that target cell-surface ectodomain proteins on both living and fixed cells.
量子点是一种新型荧光平台，有潜力成为用于标记生物探针（如抗体或配体）的传统有机染料的替代品。与典型的荧光有机染料相比，硒化镉/硫化锌核壳纳米晶体（即量子点）具有更高的光稳定性，能抵抗代谢和化学降解，无毒，且发射光谱宽、激发光谱窄。当与链霉亲和素等通用衔接分子偶联时，量子点可用于标记不同的生物素化抗体或配体，而无需针对每种抗体或配体定制量子点表面化学。在本章中，我们概述了使用链霉亲和素量子点标记生物素化抗体的方法，这些抗体靶向活细胞和固定细胞上的细胞表面胞外域蛋白。
