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通过反相液相-质谱法对无细胞蛋白合成代谢的绝对定量
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Absolute Quantification of Cell-Free Protein Synthesis Metabolism by Reversed-Phase Liquid Chromatography-Mass Spectrometry
DOI:

08:06 min

October 25, 2019

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Chapters

  • 00:05Title
  • 01:01Preparation for Aniline Tagging
  • 02:41Labeling Reaction and LC/MS Procedure
  • 06:06Results: Identification and Labeling Results of Metabolites
  • 07:02Conclusion

Summary

Automatic Translation

在这里,我们提出了一个强大的协议,以量化40个化合物参与中央碳和能量代谢在无细胞蛋白质合成反应。无细胞合成混合物用抗氧化剂衍生,使用反相液相色谱进行有效分离,然后使用异位标记的内部标准通过质谱法进行量化。

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