Laboratory of Viral Diseases, NIAID, NIH, Bethesda, MD 20892, USA.
Histochem Cell Biol. 2013 Mar;139(3):501-4. doi: 10.1007/s00418-012-1063-8. Epub 2012 Dec 11.
We previously described the ribopuromyclation method (RPM) to visualize and quantitate translating ribosomes in fixed and permeabilized cells by standard immunofluorescence. RPM is based on puromycylation of nascent chains bound to translating ribosomes followed by detection of puromycylated nascent chains with a puromycin-specific mAb. We now demonstrate that emetine optimally enhances nascent chain puromycylation, and describe a modified RPM protocol for identifying ribosome-bound nascent chains in metabolically inert permeabilized cells.
我们之前描述了核糖基化嘌呤霉素酰化法（RPM），通过标准免疫荧光来可视化和定量固定和通透细胞中的翻译核糖体。RPM 基于与翻译核糖体结合的新生肽链的嘌呤霉素酰化，然后用嘌呤霉素特异性 mAb 检测嘌呤霉素化的新生肽链。我们现在证明依米丁能最优增强新生肽链的嘌呤霉素酰化，并描述了一种改良的 RPM 方案，用于鉴定代谢惰性通透细胞中与核糖体结合的新生肽链。
