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 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.

Contents:

PART I: PROTEOMICS TECHNOLOGY

• Introduction
• Expression Proteomics

PART II: COURSE MANUAL

• Equipment, Consumables, Reagents
• Sample Preparation
• Isoelectric Focusing
• SDS Polyacrylamide Gel Electrophoresis
• Straining of the Gels
• Scanning of Gels and Image Analysis
• Fluorescence Difference Gel Electrophoresis
• Spot Excision
• Sample Destaining
• In-gel Digestion
• Microscale Desalting and Concentration of Sample
• Chemical Derivatisation of the Peptide Digest
• MS Analysis
• Calibration of the MALDI-ToF MS
• Preparing for a Database Search
• PMF Database Search Unsuccessful

Trouble Shooting

Index

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