Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), 53127 Bonn, Germany.
Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), 53127 Bonn, Germany.
STAR Protoc. 2023 Dec 15;4(4):102559. doi: 10.1016/j.xpro.2023.102559. Epub 2023 Sep 14.
Protein synthesis, or mRNA translation, is the biological process through which genetic information stored in messenger RNAs is encoded into proteins. Here, we present an optimized protocol for assessing the translation rate in mouse adult microglia and cultured bone-marrow-derived macrophages. We describe steps for isolating cells, treating them with a puromycin-analog probe, and fluorescently labeling the puromycylated-polypeptide chains. We then detail their quantification by flow cytometry or with a fluorescent plate reader. For complete details on the use and execution of this protocol, please refer to Keane et al. (2021)..
蛋白质合成,或 mRNA 翻译,是将储存在信使 RNA 中的遗传信息编码为蛋白质的生物学过程。在这里,我们提供了一个优化的方案,用于评估成年小鼠小神经胶质细胞和培养的骨髓来源的巨噬细胞中的翻译速率。我们描述了分离细胞、用嘌呤霉素类似物探针处理细胞以及荧光标记被嘌呤霉素酰化的多肽链的步骤。然后,我们详细介绍了通过流式细胞仪或荧光板读数器对其进行定量的方法。有关该方案的使用和执行的完整详细信息,请参阅 Keane 等人。(2021 年)。