This volume provides state-of-the-art methods and protocols from researchers in the field of metabolism. Chapters guide readers through methodologies measuring different aspects of lipid metabolism, quantifying the amino acids, mitochondrial and intracellular by-products, detect enzyme and kinase activities, detecting a variety of metabolic pathways, and intracellular networks including nucleotide metabolism and the metabolic response to hypoxia and cytotoxic drugs. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the…mehr
This volume provides state-of-the-art methods and protocols from researchers in the field of metabolism. Chapters guide readers through methodologies measuring different aspects of lipid metabolism, quantifying the amino acids, mitochondrial and intracellular by-products, detect enzyme and kinase activities, detecting a variety of metabolic pathways, and intracellular networks including nucleotide metabolism and the metabolic response to hypoxia and cytotoxic drugs. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Metabolic Reprogramming: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
In Vivo Tissue Lipid Uptake in Antisense Oligonucleotides (ASO) Treated Mice.- In Vivo Hepatic Triglyceride Secretion Rate in Antisense Oligonucleotides (ASO) Treated Mice.- Measurement of Fatty Acid Oxidation by High Resolution Respirometry: Special Considerations for Analysis of Skeletal and Cardiac Muscle and Adipose Tissue.- In Vivo Imaging of Bone Marrow Long Chain Fatty Acid Uptake.- Quantification and Tracing of Stable Isotope into Cysteine and Glutathione.- Targeted Quantification of Amino Acids by Dansylation.- Isolation of Mitochondria from Mouse Tissues for Functional Analysis.-Methods for Monitoring Mitochondrial Biogenesis and Turnover in Cultured Hepatocytes and Mouse Liver Using MitoTimer Reporter Assay.- Quantification of Intracellular ATP Content in Ex-Vivo GC B Cells.-UHPLC-HRMS-based Analysis of S-Hydroxymethyl-Glutathione, GSH and GSSG in Human Cells.- Quantitation of glutathione and oxidized glutathione ratios from biological matrices using LC-MS/MS.- Real-Time Monitoring of Hydrogen Peroxide Levels in Yeast and Mammalian Cells.- Tracer-Based Metabolic Analysis by NMR in Intact Perfused Human Liver Tissue.- 13C isotope labeling and mass spectrometric isotope enrichment analysis in acute brain slices.- Metabolite analyses using Nuclear Magnetic Resonance (NMR) spectroscopy in plasma of patients with prostate cancer.- Screening Kinase-Dependent Phosphorylation of Key Metabolic Reprogramming Regulators.- Measuring the Oxidation State and Enzymatic Activity of Glyceraldehyde Phosphate Dehydrogenase (GAPDH).- Quantitation of Glutamine Synthetase-1 Activity in Drosophila Melanogaster.- Fixed Cell Immunofluorescence for Quantification of Hypoxia-Induced Changes in Histone Methylation.- Metabolic Reprogramming During B Cell Differentiation.- An Integrated Methodology to Quantify the Glycolytic Stress in Plasma Cell Myeloma in Response to Cytotoxic Drugs.- Spatial Analysis of Nucleotide Metabolism: From CRISPR Knock Out Cancer Cells to MALDI Imaging of Tumors.- Assessment of Metabolic Pathways and Parameters in Extracellular Matrix-Detached Cells.- Metabolic Networks: Weighted Gene Correlation Network Analysis.
In Vivo Tissue Lipid Uptake in Antisense Oligonucleotides (ASO) Treated Mice.- In Vivo Hepatic Triglyceride Secretion Rate in Antisense Oligonucleotides (ASO) Treated Mice.- Measurement of Fatty Acid Oxidation by High Resolution Respirometry: Special Considerations for Analysis of Skeletal and Cardiac Muscle and Adipose Tissue.- In Vivo Imaging of Bone Marrow Long Chain Fatty Acid Uptake.- Quantification and Tracing of Stable Isotope into Cysteine and Glutathione.- Targeted Quantification of Amino Acids by Dansylation.- Isolation of Mitochondria from Mouse Tissues for Functional Analysis.-Methods for Monitoring Mitochondrial Biogenesis and Turnover in Cultured Hepatocytes and Mouse Liver Using MitoTimer Reporter Assay.- Quantification of Intracellular ATP Content in Ex-Vivo GC B Cells.-UHPLC-HRMS-based Analysis of S-Hydroxymethyl-Glutathione, GSH and GSSG in Human Cells.- Quantitation of glutathione and oxidized glutathione ratios from biological matrices using LC-MS/MS.- Real-Time Monitoring of Hydrogen Peroxide Levels in Yeast and Mammalian Cells.- Tracer-Based Metabolic Analysis by NMR in Intact Perfused Human Liver Tissue.- 13C isotope labeling and mass spectrometric isotope enrichment analysis in acute brain slices.- Metabolite analyses using Nuclear Magnetic Resonance (NMR) spectroscopy in plasma of patients with prostate cancer.- Screening Kinase-Dependent Phosphorylation of Key Metabolic Reprogramming Regulators.- Measuring the Oxidation State and Enzymatic Activity of Glyceraldehyde Phosphate Dehydrogenase (GAPDH).- Quantitation of Glutamine Synthetase-1 Activity in Drosophila Melanogaster.- Fixed Cell Immunofluorescence for Quantification of Hypoxia-Induced Changes in Histone Methylation.- Metabolic Reprogramming During B Cell Differentiation.- An Integrated Methodology to Quantify the Glycolytic Stress in Plasma Cell Myeloma in Response to Cytotoxic Drugs.- Spatial Analysis of Nucleotide Metabolism: From CRISPR Knock Out Cancer Cells to MALDI Imaging of Tumors.- Assessment of Metabolic Pathways and Parameters in Extracellular Matrix-Detached Cells.- Metabolic Networks: Weighted Gene Correlation Network Analysis.
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