Metabolic flux measurement

Flux will be measured using the metabolomics platforms based on absolute quantification method (isotope ratio based MS technique) by LC-MS, using heavy-isotope labelled precursors of the metabolites of interest. For example, 15N labelled cysteine, glycine and glutamate will be used to determine rates of synthesis of glutathione. 15N-labelled methionine to measure S-adenosyl methionine (and its decarboxylated form, as well as methionine cycle intermediates). 15N labelled arginine is used as precursor for ornithine, putrescine, and spermidine and trypanothione synthesis. 13C labelled glucose will be used to measure production of glucose 6-phosphate, 6-phosphogluconate and pentose-phosphate. Experiments will involve adding these precursors to medium and then following their distribution into the different metabolites under normal physiological conditions and comparing that distribution over time with cells perturbed with the different stress inducing agents and also in selected RNAi and gene knockout cells defective in various enzymes.

SEEK ID: https://fairdomhub.org/studies/104

Metabolite profiling, quantification and flux quantification

Projects: SilicoTryp

Dong-Hyun Kim

Experimentalists: Dong-Hyun Kim

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Created: 10th Dec 2012 at 17:13

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