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Table Of Contents

Introduction to electrophoretic theory
1.0 Principles of electrophoresis
1.1 Electrical parameters
1.4.2 Polyacrylamide gels
1.5 Analysis of the results
1.5.1 Detection
1.5.2 Quantification
1.6 Blotting
1.6.1 Transfer
1.6.2 Detection
1.7 Protocols in this manual
1.8 References and bibliography
Polyacrylamide gel electrophoresis
2.1 Equipment choices
2.2.1 Introduction
2.2.2 Stock solutions
2.2.3 Materials and equipment
2.2.4 Procedure
2.3 Separating proteins on denaturing mini-gels
2.3.1 Introduction
2.3.2 Materials and equipment
2.3.3 Procedure
2.4 Preparing linear pore-gradient gels
2.4.1 Introduction
2.4.2 Materials and equipment
2.4.3 Procedure
2.5 Native gel electrophoresis
2.6 Separating proteins by flatbed SDS-PAGE
2.6.1 Introduction
2.6.2 Materials and equipment
2.6.3 Procedure
2.7 Troubleshooting
2.8 References and bibliography
Isoelectric focusing of proteins
3.0 Introduction
3.1 Native isoelectric focusing
3.1.1 Solutions
3.1.2 Materials and equipment
3.1.3 Procedure
3.2 Denaturing isoelectric focusing
3.2.1 Solutions
3.2.2 Materials and equipment
3.2.3 Procedure
3.3 Isoelectric focusing using immobilized pH gradient (IPG) gels
3.3.1 Materials and equipment
3.3.2 Procedure
3.5 References and bibliography
3.4 Troubleshooting
Analysis of gels
4.1 Staining gels with Coomassie Brilliant Blue
4.1.1 Coomassie Blue staining stock solutions
4.1.2 Materials and equipment
4.1.3 Standard Coomassie Blue protocol
4.1.4 Automated Coomassie Blue staining
4.2 Silver staining
4.2.1 Silver staining stock solutions
4.2.2 Materials and equipment
4.2.3 Silver staining protocol
4.3 Gel drying and storage
4.3.1 Storage
4.3.2 Drying gels by vacuum
4.3.3 Air drying using Easy Breeze™
4.4 Documentation
4.4.1 Photography
4.4.2 Densitometer
4.4.3 Scanners and digital cameras
4.5 Estimation of protein molecular weights by SDS gel electrophoresis
4.5.1 Materials and equipment
4.5.2 Procedure
4.7 References and bibliography
4.6 Troubleshooting
Glossary
Index
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Electrophoresis Manual

Electrophoresis Manual

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Published by: Madhusudhan Narayanswamy on Jun 09, 2011
Copyright:Attribution Non-commercial

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06/09/2011

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