This volume explores the latest metabolic flux analysis (MFA) techniques that cover the analysis of cellular, organ level, and whole-body metabolism. The chapters in this book discuss topics such as deutrium tracing; isotopologue fractions using GC-TOF; non-targeted mass isotopolome analysis; large-scale profiling of cellular metabolic activities using deep 13C labeling medium; metastases in mice; SWATH; Exo-MFA; metabolic flux from time-course metabolomics; and thermodynamic approaches in flux analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, Metabolic Flux Analysis in Eukaryotic Cells: Methods and Protocols is a valuable resourcefor both experts in MFA techniques and researchers getting involved in the role of quantitative studies to uncover the dysregulated pathways in human diseases.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt
Determination of Isotopologue and Tandem Mass Isotopologue Fractions using Gas Chromatography Chemical Ionization Time of Flight Mass Spectrometry: Methodology and Uncertainty of Measurement.- Non-Targeted Mass Isotopolome Analysis using Stable Isotope Patterns to Identify Metabolic Changes.- Liquid Chromatography Methods for Separation of Polar and Charged Intracellular Metabolites for 13C Meabolic Flux Analysis.- Deuterium Tracing to Interrogate Compartment-Specific NAD(p)H Metabolism in Cultured Mammalian Cells.- Large-Scale Profiling of Cellular Metabolic Activities using Deep 13C Labeling Medium.- Analyzing the Metabolism of Metastases in Mice.- Robusy Analytical methods for the Accurate Quantification of the Total Biomass Composition of Mammalian Cells.- Quantifying Nitric Oxide Flux Distributions.- SWATH: A Data-Independent Tandem Mass Spectrometry Method to Quantify 13C Enrichment in Cellular Metabolites and Fragments.- Quantifying Metabolic Transfer Mediated by Extracellular Vesicles using Exo-MFA: An Integrated Empirical and COmputational Platform.- Comparative Metabolic Network Flux Analysis to Identify Differences in Cellular Metabolism.- Software Supporting a Workflow of Quantitative Dynamic Flux Maps Estimation in Central Metabolism from SIRM Experimental Data.- Inferring Metabolic Flux from Time-Course Metabolomics.- Using Metabolic Network Reconstructions to Predict Drug Targets and Off-Target Effects.- Single-Cell Digital Twins for Cancer Preclinical Investigation.- Supply and Demand Analysis of Autophagy.- Thermodynamic Approaches in Flux Analysis.