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Proteomics in Practice
A Laboratory Manual of Proteome Analysis
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A combined review, manual and reference for the successful analysis of proteins using the classical approach of 2-D gel electrophoresis, mass spectrometry and related sequence database inquiries.The first section, written in a textbook style, introduces the entire technology, while the second section represents a comprehensive laboratory manual spanning the full range of methods from sample preparation to protein identification. Alternative methods and procedures are only suggested for those cases where the "default" procedure would fail to deliver adequate results. The third section is a uniq...
A combined review, manual and reference for the successful analysis of proteins using the classical approach of 2-D gel electrophoresis, mass spectrometry and related sequence database inquiries.
The first section, written in a textbook style, introduces the entire technology, while the second section represents a comprehensive laboratory manual spanning the full range of methods from sample preparation to protein identification. Alternative methods and procedures are only suggested for those cases where the "default" procedure would fail to deliver adequate results. The third section is a unique troubleshooting guide, designed to answer many of the frequently asked questions regarding proteome analysis. The final section contains a thorough reference list to guide interested readers towards further detail.
Intended for all those wishing to go beyond the theoretical aspects of proteome analysis, this book is targeted at research groups within academia and industry, course instructors, research assistants and graduate students.
Special feature: Updates of recipes as well as animations illustrating crucial methodological steps are available on the Web at http://www.wiley-vch.de/books/info/3-527-30354-5
"This book says straight from the start that it will tell you how to do proteomics by 2D electrophoresis, followed by peptide mass fingerprinting, and then CAF MALDI sequence analysis" .................. ProteoMonitor
Foreword
Preface
Abbreviations, Symbols, Units
Glossary of Terms
PART I: PROTEOMICS TECHNOLOGY
Introduction
Expression Proteomics
Two-dimensional Electrophoresis
Spot Handling
Mass Spectrometry
Protein Identification by Database Searching
Methods of Proteomics
PART II: COURSE MANUAL
Step 1: Sample Preparation
Step 2: Isoelectric Focusing
Step 3: SDS Polyacrylamide Gel Electrophoresis
Step 4: Staining of the Gels
Step 5: Scanning of Gels and Image Analysis
Step 6: 2D DIGE
Step 7: Spot Excision
Step 8: Sample Destaining
Step 9: In-gel Digestion
Step 10: Microscale Purification
Step 11: Chemical Derivatisation of the Peptide Digest
Step 12: MS Analysis
Step 13: Calibration of the MALDI-ToF MS
Step 14: Preparing for a Database Search
Step 15: PMF Database Search Unsuccessful
Appendices
Trouble Shooting Guide
References
Animations of Proteomic Separations (Basic Principles)
Erratum
Addendum
The first section, written in a textbook style, introduces the entire technology, while the second section represents a comprehensive laboratory manual spanning the full range of methods from sample preparation to protein identification. Alternative methods and procedures are only suggested for those cases where the "default" procedure would fail to deliver adequate results. The third section is a unique troubleshooting guide, designed to answer many of the frequently asked questions regarding proteome analysis. The final section contains a thorough reference list to guide interested readers towards further detail.
Intended for all those wishing to go beyond the theoretical aspects of proteome analysis, this book is targeted at research groups within academia and industry, course instructors, research assistants and graduate students.
Special feature: Updates of recipes as well as animations illustrating crucial methodological steps are available on the Web at http://www.wiley-vch.de/books/info/3-527-30354-5
"This book says straight from the start that it will tell you how to do proteomics by 2D electrophoresis, followed by peptide mass fingerprinting, and then CAF MALDI sequence analysis" .................. ProteoMonitor
Foreword
Preface
Abbreviations, Symbols, Units
Glossary of Terms
PART I: PROTEOMICS TECHNOLOGY
Introduction
Expression Proteomics
Two-dimensional Electrophoresis
Spot Handling
Mass Spectrometry
Protein Identification by Database Searching
Methods of Proteomics
PART II: COURSE MANUAL
Step 1: Sample Preparation
Step 2: Isoelectric Focusing
Step 3: SDS Polyacrylamide Gel Electrophoresis
Step 4: Staining of the Gels
Step 5: Scanning of Gels and Image Analysis
Step 6: 2D DIGE
Step 7: Spot Excision
Step 8: Sample Destaining
Step 9: In-gel Digestion
Step 10: Microscale Purification
Step 11: Chemical Derivatisation of the Peptide Digest
Step 12: MS Analysis
Step 13: Calibration of the MALDI-ToF MS
Step 14: Preparing for a Database Search
Step 15: PMF Database Search Unsuccessful
Appendices
Trouble Shooting Guide
References
Animations of Proteomic Separations (Basic Principles)
Erratum
Addendum