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Author:
Date:
2014-04-20
[Abstract] Labeling of newly-synthesized polypeptides with radioactive amino acids followed by immunoprecipitation allows quantitative analysis of the fate of a given protein in a time-dependent manner. This biochemical approach is usually used to study a variety of processes, such as protein folding, co-translational modifications, intracellular transport, and even its rate of degradation. Here, I describe step by step a simple technique to both label newly-synthesized influenza A virus (IAV) hemagglutinin (HA) with [35S]-methionine and then follow its maturation and transport through the secretory pathway by SDS-PAGE and fluorography (Magadan et al., 2013).
[摘要] 用放射性氨基酸标记新合成的多肽，随后免疫沉淀允许以时间依赖性方式定量分析给定蛋白质的命运。 这种生物化学方法通常用于研究各种过程，如蛋白质折叠，共翻译修饰，细胞内转运，甚至其降解速率。 在这里，我逐步描述一个简单的技术，以标记新合成的甲型流感病毒（IAV）血凝素（HA）与[35 S] - 甲硫氨酸，然后跟随其成熟和运输通过分泌 通过SDS-PAGE和荧光成像（Magadan等人，2013）。
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