Proteomics in practice : a guide to successful experimental design / / edited by Reiner Westermeier, Tom Naven, Hans-Rudolf Höpker

• Record Nr.: UNINA9910144117403321
• Pubbl/distr/stampa: Weinheim, Germany : , : WILEY-VCH Verlag GmbH & Co. KGaA, , 2008; ©2008
• ISBN: 1-281-94680-X; 9786611946807; 3-527-62229-2; 3-527-62230-6
• Edizione: [Second, completely revised edition.]
• Descrizione fisica: 1 online resource (504 p.)
• Disciplina: 572.6; 572.64
• Soggetti: Proteomics; Proteins - Analysis; Molecular biology; Electronic books.
• Lingua di pubblicazione: Inglese
• Formato: Materiale a stampa
• Livello bibliografico: Monografia
• Note generali: Description based upon print version of record.
• Nota di bibliografia: Includes bibliographical references and index.
• Nota di contenuto: Proteomics in Practice; Contents; Preface; Foreword; Abbreviations, Symbols, Units; Introduction; 1 History; 2 Critical Points; 2.1 Challenges of the Protein Samples; 2.2 Challenges of the Analysis Systems; 3 Proteomics Strategies; 3.1 Proteome Mapping; 3.2 Differential Analysis; 3.3 Time Point Experiments; 3.4 Verification of Targets or Biomarkers; 3.5 Integration of Results into Biological Context; 3.6 Systems Biology; 4 Concept of Experimental Planning; 4.1 Biological Replicates; 4.2 Pooling of Samples: Yes or No?; 4.3 Pre-fractionation of Samples: Yes or No?; 4.4 Which is the Best Workflow to Start With?Part I: Proteomics Technology; 1 Electrophoretic Techniques; 1.1 The Principle of Electrophoresis and Some Methodological Background; 1.1.1 Free Flow Electrophoretic Methods; 1.1.2 Gels for Electrophoretic Techniques; 1.1.3 Electroendosmosis Effects; 1.2 Polyacrylamide Gel Electrophoresis; 1.2.1 The Polyacrylamide Gel; 1.2.2 SDS Polyacrylamide ; Gel Electrophoresis; 1.2.3 Blue Native Electrophoresis; 1.2.4 Cationic Detergent Electrophoresis; 1.3 Blotting; 1.3.1 Electrophoretic Transfer; 1.3.2 Protein Detection on the Membrane; 1.4 Isoelectric Focusing1.4.1 Theoretical Background; 1.4.2 Preparation of IEF Gels; 1.4.3 Isoelectric Focusing in Proteomics; 1.5 Two-dimensional Electrophoresis; 1.5.1 Sample Preparation; 1.5.2 Pre-labeling of Proteins for Difference Gel Electrophoresis; 1.5.3 First Dimension: Isoelectric Focusing in IPG Strips; 1.5.4 Second Dimension: SDS Electrophoresis; 1.5.5 Detection of Protein Spots; 1.6 Image Analysis; 1.6.1 Image Acquisition; 1.6.2 Image Analysis and Evaluation; 1.6.3 Use of 2-D Electrophoresis Data; 1.7 Spot Handling; 1.7.1 Spot Picking; 1.7.2 Protein Cleavage; 2 Liquid Chromatography Techniques2.1 Basic Principles of Important Liquid Chromatography Techniques; 2.1.1 Ion Exchange Chromatography; 2.1.2 Reversed Phase Chromatography; 2.1.3 Affinity Chromatography; 2.1.4 Gel Filtration; 2.2 Strategic Approach and General Applicability; 2.3 Liquid Chromatography Techniques and Applications in Proteome Analysis; 2.3.1 Peptide Separation; 2.3.2 2DLC Peptide Separation; 2.3.3 Affinity Chromatography and LC-MS/MS; 2.3.4 Protein Pre-fractionation; 2.4 Practical Considerations and Application of LC-based Protein Pre-fractionation; 2.4.1 Sample Extraction and Preparation2.4.2 Experimental Setup; 2.4.3 Ion Exchange Chromatography and Protein Pre-fractionation; 2.4.4 Reversed Phase Chromatography and Protein Pre-fractionation; 2.4.5 Fraction Size and Number of Fractions; 2.5 Critical Review and Outlook; 3 Mass Spectrometry; 3.1 Ionization; 3.1.1 Matrix Assisted Laser Desorption Ionization; 3.1.2 Electrospray Ionization; 3.2 Ion Separation; 3.2.1 Time-of-Flight Analyzer; 3.2.2 Triple Quadrupole Analyzer; 3.2.3 Quadrupole Ion Trap; 3.2.4 Quadrupole Time-of-Flight; 3.2.5 Hybrid Triple Quadrupole Linear Ion Trap; 3.2.6 TOF/TOF Analyzer
• Sommario/riassunto: Still the only concise practical guide to laboratory experiments in proteomics, this new edition now also covers DIGE technology and liquid-chromatography, while the troubleshooting section has been considerably extended. Adopting a practical approach, the authors present the relevant techniques and explain the route to successful experimental design and optimal method selection. They cover such electrophoretic techniques as isoelectric focusing, SDS page, 2-D page, and DIGE, as well as liquid-chromatography techniques, such as ion exchange, affinity chromatography and reversed-phase HPLC.