Protocol for detecting glycoRNAs using metabolic labeling and northwestern blot

Autor: Luoyi Li, Ningning Zhang, Carla Pantoja, Yadong Wang, Jun Lu
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: STAR Protocols, Vol 5, Iss 4, Pp 103321- (2024)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2024.103321
Popis: Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Databáze: Directory of Open Access Journals