Protocol for quantifying N-Myc and global protein translation in neuroblastoma cells using click chemistry on polyvinylidene fluoride membranes

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