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by Reiner Westermeier and Tom Naven
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 .
Hardback
Pages: 342
Published: 2002
Contents
Preface
Foreword
Abbreviations, Symbols, Units
Glossary of Terms
PART I: PROTEOMICS TECHNOLOGY
Introduction
Expression Proteomics
PART II: COURSE MANUAL
Equipment, Consumables, Reagents
Step 1: Sample Preparation
Step 2: Isoelectric Focusing
Step 3: SDS Polyacrylamide Gel Electrophoresis
Step 4: Straining of the Gels
Step 5: Scanning of Gels and Image Analysis
Step 6: Fluorescence Difference Gel Electrophoresis
Step 7: Spot Excision
Step 8: Sample Destaining
Step 9: In-gel Digestion
Step 10: Microscale Desalting and Concentration of Sample
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
Trouble Shooting
References
Index
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