一种快速、廉价、可靠的protocol，用于免疫荧光检测 2D 和 3D 集落中的多能和分化的小鼠胚胎干细胞。
A quick, cheap, and reliable protocol for immunofluorescence of pluripotent and differentiating mouse embryonic stem cells in 2D and 3D colonies.
机构信息
Department of Cell Biology, Neurobiology and Biophysics, Utrecht University, 3584 CH Utrecht, the Netherlands.
Department of Cell Biology, Neurobiology and Biophysics, Utrecht University, 3584 CH Utrecht, the Netherlands.
出版信息
STAR Protoc. 2023 Mar 17;4(1):102000. doi: 10.1016/j.xpro.2022.102000. Epub 2023 Jan 20.
Immunofluorescent labeling is a widely used method to visualize endogenous proteins. It can be expensive and difficult to stain mouse embryonic stem cells (mESCs) because they require expensive growth media, prefer specific substrates, grow in 3D, and have loose cell-substrate adhesion. Here we propose a half-a-day, cheap, easy-to-follow, and reproducible protocol for immunofluorescence of mESCs. This protocol has been streamlined to allow a fast visualization of the investigated proteins, and we provide tips specific to stem cell culture. For complete details on the use and execution of this protocol, please refer to Chaigne et al. (2021)..
免疫荧光标记是一种广泛用于可视化内源性蛋白质的方法。然而，由于其昂贵的生长培养基需求、对特定底物的偏好、三维生长方式以及松散的细胞-基质附着特性，对小鼠胚胎干细胞（mESCs）进行免疫荧光染色既昂贵又困难。在这里，我们提出了一种耗时半天、廉价、易于遵循且可重复的 mESC 免疫荧光染色方案。该方案已被精简，以允许快速可视化研究蛋白，并提供了针对干细胞培养的具体技巧。如需详细了解该方案的使用和执行，请参考 Chaigne 等人（2021 年）的研究。
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