Metabolic Flux Analysis in Eukaryotic Cells: Methods and Protocols

✍ Scribed by Deepak Nagrath

• Publisher: Springer US;Humana
• Year: 2020
• Tongue: English
• Leaves: 377
• Series: Methods in Molecular Biology 2088
• Edition: 1st ed. 2020
• Category: Library

✦ Synopsis

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 resource for both experts in MFA techniques and researchers getting involved in the role of quantitative studies to uncover the dysregulated pathways in human diseases.

✦ Table of Contents

Front Matter ....Pages i-xiii
Determination of Isotopologue and Tandem Mass Isotopologue Ratios Using Gas Chromatography Chemical Ionization Time of Flight Mass Spectrometry - Methodology and Uncertainty of Measurement (Teresa Mairinger, Stephen Hann)....Pages 1-16
Non-Targeted Mass Isotopolome Analysis Using Stable Isotope Patterns to Identify Metabolic Changes (Christian-Alexander Dudek, Lisa Schlicker, Karsten Hiller)....Pages 17-32
Liquid Chromatography Methods for Separation of Polar and Charged Intracellular Metabolites for 13C Metabolic Flux Analysis (Damini Jaiswal, Anjali Mittal, Deepak Nagrath, Pramod P. Wangikar)....Pages 33-50
Deuterium Tracing to Interrogate Compartment-Specific NAD(P)H Metabolism in Cultured Mammalian Cells (Esther W. Lim, Seth J. Parker, Christian M. Metallo)....Pages 51-71
Large-Scale Profiling of Cellular Metabolic Activities Using Deep 13C Labeling Medium (Nina Grankvist, Jeramie D. Watrous, Mohit Jain, Roland Nilsson)....Pages 73-92
Analyzing the Metabolism of Metastases in Mice (Patricia Altea-Manzano, Dorien Broekaert, João A. G. Duarte, Juan Fernández-García, Mélanie Planque, Sarah-Maria Fendt)....Pages 93-118
Robust Analytical Methods for the Accurate Quantification of the Total Biomass Composition of Mammalian Cells (Diana Széliová, Harald Schoeny, Špela Knez, Christina Troyer, Cristina Coman, Evelyn Rampler et al.)....Pages 119-160
Quantifying Nitric Oxide Flux Distributions (Darshan M. Sivaloganathan, Xuanqing Wan, Mark P. Brynildsen)....Pages 161-188
SWATH: A Data-Independent Tandem Mass Spectrometry Method to Quantify 13C Enrichment in Cellular Metabolites and Fragments (Damini Jaiswal, Pramod P. Wangikar)....Pages 189-204
Quantifying Metabolic Transfer Mediated by Extracellular Vesicles Using Exo-MFA: An Integrated Empirical and Computational Platform (Abhinav Achreja, Noah Meurs, Deepak Nagrath)....Pages 205-221
Comparative Metabolic Network Flux Analysis to Identify Differences in Cellular Metabolism (Sarah McGarrity, Sigurður T. Karvelsson, Ólafur E. Sigurjónsson, Óttar Rolfsson)....Pages 223-269
Software Supporting a Workflow of Quantitative Dynamic Flux Maps Estimation in Central Metabolism from SIRM Experimental Data (Vitaly A. Selivanov, Silvia Marin, Josep Tarragó-Celada, Andrew N. Lane, Richard M. Higashi, Teresa W.-M. Fan et al.)....Pages 271-298
Inferring Metabolic Flux from Time-Course Metabolomics (Scott Campit, Sriram Chandrasekaran)....Pages 299-313
Metabolic Network Reconstructions to Predict Drug Targets and Off-Target Effects (Kristopher Rawls, Bonnie V. Dougherty, Jason Papin)....Pages 315-330
Single-cell Digital Twins for Cancer Preclinical Investigation (Marzia Di Filippo, Chiara Damiani, Marco Vanoni, Davide Maspero, Giancarlo Mauri, Lilia Alberghina et al.)....Pages 331-343
Supply and Demand Analysis of Autophagy (André du Toit, Ben Loos, Jan Hendrik S. Hofmeyr)....Pages 345-357
Thermodynamic Approaches in Flux Analysis (Sabine Peres, Vincent Fromion)....Pages 359-367
Back Matter ....Pages 369-370

✦ Subjects

Life Sciences; Metabolomics; Cell Biology; Biotechnology