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Vloeistofchromatografie gekoppeld aan brekingsindex of massaspectrometrische detectie voor metabolietprofilering in op lysaat gebaseerde celvrije systemen
JoVE Journal
Bioengineering
This content is Free Access.
JoVE Journal Bioengineering
Liquid Chromatography Coupled to Refractive Index or Mass Spectrometric Detection for Metabolite Profiling in Lysate-based Cell-free Systems
DOI:

14:42 min

September 23, 2021

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Chapters

  • 00:04Introduction
  • 01:08Starting, Stopping, and Processing Time Course CFME Reactions for HPLC-RID Quantification
  • 03:18Creating a Sequence Table for Autosampling and Start the HPLC-RID System for Data Acquisition
  • 04:54Extracting and Analyzing Data Post-Run
  • 07:20Starting, Stopping, and Processing Time Course Isotope Tracing CFME Reactions for LC-MS/MS Quantification
  • 09:20Setting Up a Run Sequence and Starting the LC-MS/MS Run
  • 10:16Calculating Negative Mode Masses of 13C-Labeled Glucose-Derived Metabolites and Searching for the m/z Features of These Analytes in Filtered Data
  • 11:45Results: Metabolite Profiling in Lysate-based Cell-free Systems
  • 13:54Conclusion

Summary

Automatic Translation

De protocollen beschrijven hoogwaardige vloeistofchromatografiemethoden gekoppeld aan brekingsindex of massaspectrometrische detectie voor het bestuderen van metabole reacties in complexe op lysaat gebaseerde celvrije systemen.

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