/
/
Absolute Quantification of Cell-Free Protein Synthesis Metabolism by Reversed-Phase Liquid Chromatography-Mass Spectrometry
A subscription to JoVE is required to view this content. Sign in or start your free trial.
8,855 Views
•
08:06 min
•
October 25, 2019
DOI:
Summary
Automatically generated
Here, we present a robust protocol to quantify 40 compounds involved in central carbon and energy metabolism in cell-free protein synthesis reactions. The cell-free synthesis mixture is derivatized with aniline for effective separation using reversed-phase liquid chromatography and then quantified by mass spectrometry using isotopically labelled internal standards.
Read Article
Cite this Article
Vilkhovoy, M., Dai, D., Vadhin, S., Adhikari, A., Varner, J. D. Absolute Quantification of Cell-Free Protein Synthesis Metabolism by Reversed-Phase Liquid Chromatography-Mass Spectrometry. J. Vis. Exp. (152), e60329, doi:10.3791/60329 (2019).
Copy