Methodologies for Metabolomics : Experimental Strategies and Techniques


Norbert. Lutz
Bok Engelsk 2013 · Electronic books.
Medvirkende
Utgitt
Cambridge : Cambridge University Press , 2013
Omfang
1 online resource (642 p.)
Opplysninger
Description based upon print version of record.. - Contents; Contributors; Section 1 Basic Methodological Strategies in Metabolomic Research; 1 Exploring the Human Metabolome by Nuclear Magnetic Resonance Spectroscopy and Mass Spectrometry; 1. Brief History of Human Metabolomics; 2. Sample Preparation for Human Metabolomic Studies; 3. Human Metabolite Identification by Nuclear Magnetic Resonance Spectroscopy; 4. Human Metabolite Identification by Gas Chromatography-Mass Spectrometry; 5. Human Metabolite Identification by Liquid Chromatography-Mass Spectrometry; 6. Comparison of Technologies; 7. Conclusion; Acknowledgments. - 1.2. Gene Transduction: Viral Vectors2. Gene Silencing as a Tool for Metabolomic Studies; 3. Validation of Gene Overexpression or Silencing by mRNA; 3.1. Gene Expression Analysis; 3.1.1. Reverse Transcriptase Polymerase Chain Reaction; 3.1.2. Real-Time Polymerase Chain Reaction; 4. Validation of Gene Overexpression or Silencing by Protein Level Analysis and Enzyme Assays; 4.1. Western Blotting; 4.1.1. Blotting; 4.1.2. Immunodetection; 4.2. Enzyme Assays; 4.2.1. Protocols for Analysis of Choline Kinase- Activity; 4.2.2. Protocol for Analysis of Phospholipase D Activity. - 2 Methodological Requirements for Lipidomics Research1. Sample Preparation and Lipid Extraction; 2. Sample Preparation; 3. Lipid Extraction; 4. Sample Preparation and Lipid Extraction of Subcellular Organelles; 5. Chromatography-Coupled Mass Spectrometry of Lipids; 6. Shotgun Lipidomics - Direct Infusion-Based Mass Spectrometry of Lipids; 7. Postacquisition Data Processing and Bioinformatics; 8. Conclusion; Acknowledgments; 3 Biological Methods for Metabolic Research; 1. Gene Overexpression as a Tool for Metabolomic Studies; 1.1. Gene Transfection: Plasmid Vectors. - 2. Considerations in the Growth, Harvest, Preparation, and Extraction of Plant Materials. - 4 Considerations in Sample Preparation, Collection, and Extraction Approaches Applied in Microbial, Plant, and Mammalian Metabolic Profiling1. Considerations in the Culturing, Collection, Preparation, and Extraction of Microbial Samples; 1.1. Sample Types; 1.2. Culture Methods; 1.2.1. Batch Growth; 1.2.2. Continuous Culture; 1.2.3. Selection of Media; 1.3. Experimental Considerations in the Collection and Preparation of Samples; 1.3.1. Quenching of Cultures; 1.3.2. Assessment of Leakage; 1.3.3. Extraction of Intracellular Metabolites. - 5. Enzymatic Assays as a Tool of Phospholipid Metabolism Analysis: Effects of Choline Kinase- Inhibition on the Balance of Phospholipid Metabolites5.1. Protocol for Lipid Analysis; 5.2. Protocol for Analysis of Phosphatidylinositol-3-Kinase Activity; 6. Phospholipid Profile Analysis by Mass Spectrometry; 6.1. Phospholipid Extraction; 6.2. Analytical Technique for Phospholipids: Mass Spectrometry; Section 2 Metabolomic Mass Spectrometry: Experimental Techniques and Bioinformatics. - Methodologies for Metabolomics provides comprehensive descriptions of the newest methodological strategies and techniques in metabolomic research.
Emner
Sjanger
Dewey
543.65 . - 543
ISBN
9780521765909

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