Electrophoresis in Practice

Inhaltsverzeichnis

Foreword.
Preface.

Abbreviations.

Part I Fundamentals.

Introduction.

1 Electrophoresis.

1.0 General.

1.1 Electrophoresis in non-restrictive gels.

1.2 Electrophoresis in restrictive gels.

2 Isotachophoresis.

2.1 Migration with the same speed.

2.2 "Ion train" separation.

2.3 Zone sharpening effect.

2.4 Concentration regulation effect.

3 Isoelectric focusing.

3.1 Principles.

3.2 Gels for IEF.

3.3 Temperature.

3.4 Controlling the pH gradient.

3.5 The kinds of pH gradients.

3.6 Protein detection in IEFgels.

3.7 Preparative isoelectric focusing.

3.8 Titration curve analysis.

4 Blotting.

4.1 Principle.

4.2 Transfer methods.

4.3 Blotting membranes.

4.4 Buffers for electrophoretic transfers.

4.5 General staining.

4.6 Blocking.

4.7 Specific detection.

4.8 Protein sequencing.

4.9 Transfer problems.

5 Interpretation of electropherograms.

5.1 Introduction.

5.2 Image analysis.

6 Proteome Analysis.

6.1 General.

6.2 Sample preparation.

6.3 Two-dimensional electrophoresis.

6.4 Detection techniques.

6.5 Image analysis.

6.6 Protein spot identification.

6.7 Bioinformatics.

6.8 Functional proteomics.

7 Instrumentation.

7.1 Current and voltage conditions.

7.2 Power supply.

7.3 Separation chambers.

7.4 Staining apparatus for gels and blots.

7.5 Automated electrophoresis.

7.6 Instruments for 2-D electrophoresis.

7.7 Safety measures.

7.8 Environmental aspects.

Part II Equipment and Methods.

Equipment for Part II.

Instrumentation.

Special laboratory equipment.

Consumables.

Chemicals.

Method 1: PAGE of dyes.

1 Sample preparation.

2 Stock solutions.

3 Preparing the casting cassette.

4 Casting ultrathin-layer gels.

5 Electrophoretic separation.

Method 2: Agarose and immuno electrophoresis.

1 Sample preparation.

2 Stock solutions.

3 Preparing the gels.

4 Electrophoresis.

5 Protein detection.

Method 3: Titration curve analysis.

1 Sample preparation.

2 Stock solutions.

3 Preparing the blank gels.

4 Titration curve analysis.

5 Coomassie and silver staining.

6 Interpreting the curves.

Method 4: Native PAGE in amphoteric buffers.

1 Sample preparation.

2 Stock solutions.

3 Preparing the empty gels.

4 Electrophoresis.

5 Coomassie and silver staining.

Method 5: Agarose IEF.

1 Sample preparation.

2 Preparing the agarose gel.

3 Isoelectric focusing.

4 Protein detection.

Method 6: PAGIEF in rehydrated gels.

1 Sample preparation.

2 Stock solutions.

3 Preparing the blank gels.

4 Isoelectric focusing.

5 Coomassie and silver staining.

6 Perspectives.

Method 7: Horizontal SDS-PAGE.

1 Sample preparation.

2 Stock solutions for gel preparation.

3 Preparing the casting cassette.

4 Gradient gel.

5 Electrophoresis.

6 Protein detection.

7 Blotting.

8 Perspectives.

Method 8: Vertical PAGE.

1 Sample preparation.

2 Stock solutions.

3 Single gel casting.

4 Multiple gel casting.

5 Electrophoresis.

6 SDS electrophoresis of small peptides.

7 Two-dimensional electrophoresis.

8 DNA electrophoresis.

9 Long shelflife gels.

10 Detection of bands.

Method 9: Semi-dry blotting of proteins.

1 Transfer buffers.

2 Technical procedure.

3 Staining of blotting membranes.

Method 10: IEF in immobilized pH gradients.

1 Sample preparation.

2 Stock solutions.

3 Immobiline recipes.

4 Preparing the casting cassette.

5 Preparing the pH gradient gels.

6 Isoelectric focusing.

7 Staining.

8 Strategies for IPG focusing.

Method 11: High-resolution 2-D electrophoresis.

1 Sample preparation.

2 Stock solutions.

3 Preparing the gels.

4 Separation conditions.

5 Staining procedures.

Method 12: PAGE of double stranded DNA.

1 Stock solutions.

2 Preparing the gels.

3 Sample preparation.

4 Electrophoresis.

5
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Electrophoresis in Practice

A Guide to Methods and Applications of DNA and Protein Separations

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Beschreibung

Details

Einband

Gebundene Ausgabe

Erscheinungsdatum

01.12.2004

Verlag

Wiley-Vch

Seitenzahl

406

Maße (L/B/H)

24/17/17,5 cm

Beschreibung

Rezension

From the reviews of previous editions:
"The illustrations are informative and to the point...The book clearly illustrates the competence of the author. He generously shares his experience and knowledge with the reader, especially in the field of gel-based electrophoresis...an excellent reference to help the reader to become acquainted with the field of electrophoresis. The book contains theoretical and ample practical information that serves not only the practician in the electrophoresis laboratory, but also students and lecturers in electrophoretic methods."
Clinical Chemistry
"The rigorous description of each method along with the extensive figures will easily allow the novice to reproduce these methods in the laboratory"
The Analyst
"An excellent book which we recommend greatly and which has not to be missed in any laboratory of cellular and molecular biology which respects itself."
Cellular and Molecular Biology
"As a comprehensive guide to the huge variety of electrophoretic methods now available, this is very good value. The superb troubleshooting appendix almost justifies the price on its own."
Laboratory Equipment Digest

Details

Einband

Gebundene Ausgabe

Erscheinungsdatum

01.12.2004

Verlag

Wiley-Vch

Seitenzahl

406

Maße (L/B/H)

24/17/17,5 cm

Gewicht

944 g

Auflage

4. Auflage

Sprache

Deutsch, Englisch

ISBN

978-3-527-31181-1

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  • Electrophoresis in Practice
  • Foreword.
    Preface.

    Abbreviations.

    Part I Fundamentals.

    Introduction.

    1 Electrophoresis.

    1.0 General.

    1.1 Electrophoresis in non-restrictive gels.

    1.2 Electrophoresis in restrictive gels.

    2 Isotachophoresis.

    2.1 Migration with the same speed.

    2.2 "Ion train" separation.

    2.3 Zone sharpening effect.

    2.4 Concentration regulation effect.

    3 Isoelectric focusing.

    3.1 Principles.

    3.2 Gels for IEF.

    3.3 Temperature.

    3.4 Controlling the pH gradient.

    3.5 The kinds of pH gradients.

    3.6 Protein detection in IEFgels.

    3.7 Preparative isoelectric focusing.

    3.8 Titration curve analysis.

    4 Blotting.

    4.1 Principle.

    4.2 Transfer methods.

    4.3 Blotting membranes.

    4.4 Buffers for electrophoretic transfers.

    4.5 General staining.

    4.6 Blocking.

    4.7 Specific detection.

    4.8 Protein sequencing.

    4.9 Transfer problems.

    5 Interpretation of electropherograms.

    5.1 Introduction.

    5.2 Image analysis.

    6 Proteome Analysis.

    6.1 General.

    6.2 Sample preparation.

    6.3 Two-dimensional electrophoresis.

    6.4 Detection techniques.

    6.5 Image analysis.

    6.6 Protein spot identification.

    6.7 Bioinformatics.

    6.8 Functional proteomics.

    7 Instrumentation.

    7.1 Current and voltage conditions.

    7.2 Power supply.

    7.3 Separation chambers.

    7.4 Staining apparatus for gels and blots.

    7.5 Automated electrophoresis.

    7.6 Instruments for 2-D electrophoresis.

    7.7 Safety measures.

    7.8 Environmental aspects.

    Part II Equipment and Methods.

    Equipment for Part II.

    Instrumentation.

    Special laboratory equipment.

    Consumables.

    Chemicals.

    Method 1: PAGE of dyes.

    1 Sample preparation.

    2 Stock solutions.

    3 Preparing the casting cassette.

    4 Casting ultrathin-layer gels.

    5 Electrophoretic separation.

    Method 2: Agarose and immuno electrophoresis.

    1 Sample preparation.

    2 Stock solutions.

    3 Preparing the gels.

    4 Electrophoresis.

    5 Protein detection.

    Method 3: Titration curve analysis.

    1 Sample preparation.

    2 Stock solutions.

    3 Preparing the blank gels.

    4 Titration curve analysis.

    5 Coomassie and silver staining.

    6 Interpreting the curves.

    Method 4: Native PAGE in amphoteric buffers.

    1 Sample preparation.

    2 Stock solutions.

    3 Preparing the empty gels.

    4 Electrophoresis.

    5 Coomassie and silver staining.

    Method 5: Agarose IEF.

    1 Sample preparation.

    2 Preparing the agarose gel.

    3 Isoelectric focusing.

    4 Protein detection.

    Method 6: PAGIEF in rehydrated gels.

    1 Sample preparation.

    2 Stock solutions.

    3 Preparing the blank gels.

    4 Isoelectric focusing.

    5 Coomassie and silver staining.

    6 Perspectives.

    Method 7: Horizontal SDS-PAGE.

    1 Sample preparation.

    2 Stock solutions for gel preparation.

    3 Preparing the casting cassette.

    4 Gradient gel.

    5 Electrophoresis.

    6 Protein detection.

    7 Blotting.

    8 Perspectives.

    Method 8: Vertical PAGE.

    1 Sample preparation.

    2 Stock solutions.

    3 Single gel casting.

    4 Multiple gel casting.

    5 Electrophoresis.

    6 SDS electrophoresis of small peptides.

    7 Two-dimensional electrophoresis.

    8 DNA electrophoresis.

    9 Long shelflife gels.

    10 Detection of bands.

    Method 9: Semi-dry blotting of proteins.

    1 Transfer buffers.

    2 Technical procedure.

    3 Staining of blotting membranes.

    Method 10: IEF in immobilized pH gradients.

    1 Sample preparation.

    2 Stock solutions.

    3 Immobiline recipes.

    4 Preparing the casting cassette.

    5 Preparing the pH gradient gels.

    6 Isoelectric focusing.

    7 Staining.

    8 Strategies for IPG focusing.

    Method 11: High-resolution 2-D electrophoresis.

    1 Sample preparation.

    2 Stock solutions.

    3 Preparing the gels.

    4 Separation conditions.

    5 Staining procedures.

    Method 12: PAGE of double stranded DNA.

    1 Stock solutions.

    2 Preparing the gels.

    3 Sample preparation.

    4 Electrophoresis.

    5