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Quantification of Stable Isotope Label in Metabolites via Mass Spectrometry

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1056)

Abstract

Isotope labelling experiments with stable or radioactive isotopes have long been an integral part of biological and medical research. Labelling experiments led to the discovery of new metabolic pathways and made it possible to calculate the fluxes responsible for a metabolic phenotype, i.e., the qualitative and quantitative composition of metabolites in a biological system. Prerequisite for efficient isotope labelling experiments is a reliable and precise method to analyze the redistribution of isotope label in a metabolic network. Here we describe the use of the CORRECTOR program, which utilizes matrix calculations to correct mass spectral data from stable isotope labelling experiments for the distorting effect of naturally occurring stable isotopes (NOIs). CORRECTOR facilitates and speeds up the routine quantification of experimentally introduced isotope label from multiple mass spectral readouts, which are generated by routine metabolite profiling when combined with stable isotope labelling experiments.

Key words

Stable isotope labelling Stable isotope tracing Flux CORRECTOR software Metabolite profiling 

Notes

Acknowledgements

This work was supported by the Max Planck society and a grant from the DFG (Deutsche Forschungsgemeinschaft) to JK. The CORRECTOR software tool was supported in part by the European META-PHOR project, FOOD-CT-2006-036220, and the GoFORSYS project (http://www.goforsys.de/) funded by the Bundesministerium für Bildung und Forschung (BMBF). JH and BH were supported by BMBF grants 0315295 and 0315426A.

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  1. 1.Systems Biology Research Group, Department of Physiology and Cell BiologyLeibniz Institute of Plant Genetics and Crop Plant Research (IPK)GaterslebenGermany
  2. 2.Department of Theory, Max-Planck-Institute of Coal ResearchMülheim an der RuhrRuhrGermany
  3. 3.Department of Molecular PhysiologyMax-Planck-Institute of Molecular Plant Physiology (MPIMP)Potsdam-GolmGermany

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