Li Luoyi, Zhang Ningning, Pantoja Carla, Wang Yadong, Lu Jun
Yale Stem Cell Center, Yale University School of Medicine, New Haven, CT 06520, USA; Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA.
Yale Stem Cell Center, Yale University School of Medicine, New Haven, CT 06520, USA; Department of Genetics, Yale University School of Medicine, New Haven, CT 06510, USA; Yale Center for RNA Science and Medicine, New Haven, CT 06520, USA.
STAR Protoc. 2024 Dec 20;5(4):103321. doi: 10.1016/j.xpro.2024.103321. Epub 2024 Sep 19.
GlycoRNAs are glycosylated RNAs that can be detected in many cell types and often partly reside on the outer cell surface, with a recently demonstrated role in mediating neutrophil-endothelium interaction. Here, we present a protocol for glycoRNA detection based on metabolic tracing and northwestern blot. We describe steps for metabolic labeling of cells, extraction and purification of RNA, biotin labeling of RNA, and northwestern blot for glycoRNA detection. We also incorporate optimized conditions for biotin labeling, RNA dye, and membrane blocking. For complete details on the use and execution of this protocol, please refer to Zhang et al..
糖基化RNA（GlycoRNAs）是一类可在多种细胞类型中检测到的糖基化RNA，通常部分位于细胞外表面，最近已证明其在介导中性粒细胞与内皮细胞相互作用中发挥作用。在此，我们介绍一种基于代谢追踪和蛋白质印迹杂交法（northwestern blot）的糖基化RNA检测方法。我们描述了细胞代谢标记、RNA提取与纯化、RNA生物素标记以及用于糖基化RNA检测的蛋白质印迹杂交法的步骤。我们还纳入了生物素标记、RNA染料和膜封闭的优化条件。有关本方法的使用和实施的完整详细信息，请参考Zhang等人的文献。
