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Updated: Jun 12, 2025

Quantitative Immunofluorescence to Measure Global Localized Translation
Published on: August 22, 2017
Protocol for quantifying N-Myc and global protein translation in neuroblastoma cells using click chemistry on
Pamorn Chittavanich1, Duangporn Saengwimol2, Ariestya Indah Permata Sari1
1Program in Translational Medicine, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand.
Abstract:
MYCN amplification is a hallmark of aggressive neuroblastoma, driving N-Myc overexpression and enhancing protein synthesis, making these processes potential therapeutic targets. Here, we present a protocol for quantifying nascent N-Myc and global protein translation in neuroblastoma cells. This protocol describes the steps for labeling nascent proteins and performing an optimized click chemistry reaction directly on the membrane after blotting, enabling high-sensitivity detection and analysis. Adaptable to other proteins of interest, this approach provides valuable insights into neuroblastoma protein synthesis. For complete details on the use and execution of this protocol, please refer to Chittavanich et al.1.

