A flow-cytometry-based assessment of global protein synthesis in human senescent cells

Senescent cells constantly experience stressful conditions and restrain their protein translation to cope with it. Here, we present a detailed protocol to measure the rate of global protein synthesis using L-azidohomoalanine (L-AHA)-based click chemistry in human senescent fibroblasts. We optimized...

Descripción completa

Detalles Bibliográficos
Autores principales: Lee, Yeonghyeon, Kim, Jaejin, Jeon, Taerang, Roh, Kyeonghwan, Kim, Mi-Sung, Kang, Chanhee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8450257/
https://www.ncbi.nlm.nih.gov/pubmed/34585148
http://dx.doi.org/10.1016/j.xpro.2021.100809
Descripción
Sumario:Senescent cells constantly experience stressful conditions and restrain their protein translation to cope with it. Here, we present a detailed protocol to measure the rate of global protein synthesis using L-azidohomoalanine (L-AHA)-based click chemistry in human senescent fibroblasts. We optimized several aspects of the procedure, including senescence induction, a flow cytometry analysis of senescent cells, and the duration of L-AHA incorporation. This protocol uses senescent human fibroblasts but can be applied to other types of cells or circumstances. For complete details on the use and execution of this protocol, please refer to Lee et al. (2021).

Ejemplares similares