Centrifugation in Density Gradients - 1st Edition - ISBN: 9780125645805, 9781483282060

Centrifugation in Density Gradients

1st Edition

Authors: C. A. Price
eBook ISBN: 9781483282060
Imprint: Academic Press
Published Date: 28th January 1982
Page Count: 446
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Description

Centrifugation in Density Gradients provides information pertinent to the fundamental aspects of density gradient centrifugation. This book discusses the benefits of density gradient centrifugation to membrane-bound particles.

Organized into nine chapters, this book begins with an overview of the method of differential or fractional centrifugation. This text then explores the physical basis of density gradient centrifugation. Other chapters deal with the nuts and bolts of density gradient centrifugation, the construction and composition of gradients, the properties and operation of centrifuge systems, and certain arcane but highly useful procedures. This book discusses as well density gradient centrifugation in the analytical ultracentrifuge. The final chapter deals with a collection of protocols for separating particles ranging in size from whole cells to macromolecules.

This book is intended to be suitable for readers who need to separate biological particles. Biologists, chemists, biochemists, cytologists, physiologists, scientists, and research workers will also find this book useful.

Table of Contents


Foreword

Preface

1 Particle Abstraction in Biology—An Introduction

Text

2 Origins of Density Gradient Centrifugation

Text

3 Sedimentation Theory: A Semiquantitative Approach

3.1 Behavior of Particles in Centrifugal Fields

3.2 Rate Separations

3.3 Equilibrium Density. Isopycnic Separations and Sedimentation Equilibrium

References

4 Sedimentation Theory: A More Rigorous Approach

4.1 Physical Description

4.2 Equation of Motion of a Single Particle in a Centrifugal Field

4.3 Motion of Dilute Collections of Particles in a Centrifugal Field

4.4 Motion of Boundaries and Zones

References

5 Gradient Materials and Construction of Gradients

5.1 Choice of Gradient Materials

5.2 Concentration Conventions and Stock Solutions

5.3 Gradient Generators

5.4 Density Measurements

References

6 Techniques of Preparative Density Gradient Centrifugation

6.1 Planning a Separation

6.2 Swinging-Bucket Separations

6.3 Zonal Separations

Data Processing

References

7 Special Topics

7.1 Continuous-Flow Centrifugation with Isopycnic Banding

7.2 Flotation

7.3 Gradient Centrifugation in Angle and Vertical Rotors

7.4 Multiple-Alternate-Channel-Selection (MACS)

7.5 Reorienting Gradient Centrifugation

7.6 Seals: Polishing and Maintenance

7.7 Estimation of Sedimentation Coefficients

7.8 Transparent Rotors

7.9 Environmental Gradients

7.10 Sterilization

7.11 Other Uses of Density-Gradient Centrifugation

References

8 Analytical Centrifugation in Density Gradients

8.1 The Hardware of Analytical Centrifugation

8.2 Analytical Band Sedimentation in Density Gradients

8.3 Sedimentation-Diffusion Equilibrium in a Density Gradient

References

9 Protocols for the Separation and Analysis of Some Specific Particles

9.1 Whole Organisms

9.2 Whole Cells

9.3 Membrane-Bound Organelles

9.4 Multimolecular Systems

9.5 Macromolecules

References

Appendix A: Some Useful Units, Values, and Conversions

A1. Mathematical Values and Relations

A2. Physical and Chemical Values and Conversions

A3. Values of ω2t for Different Speeds and Times

A4. Relation of ω2t to RPM (Fig. A2)

A5. Nomograph Relating Centrifugal Field, Radius, and Speed (Fig. A3)

Appendix B: Properties of Particles

B1. Natural Particles

B2. Frictional Coefficients for Different Shapes

B3. Artificial Particles

Appendix C: Properties of Gradient Materials

C1. Density and Viscosity of Aqueous Solutions of Sucrose

C2. Density and Viscosity of Sucrose As Functions of Concentration

C3. Refractive Index of Sucrose Solutions

C4. Recipes for Preparing Stock Solutions of Aqueous Sucrose

C5. Dilution of Stock Solutions of Sucrose

C6. Coefficients for Density and Refractive Index of Alkali Salts

C7. Additive Mixing Relations for Dilutions of CsCI Solutions

C8. Density at 25°C of CsCI Solutions as a Function of Refractive Index

C9. Properties of Sorbitol

C10. Density, Refractive Index, and Viscosity of Ficoll

C11. Properties of Colloidal Silicas

C12. Density, Refractive Index, and Viscosity of Metrizamide

C13. Radioopaque Agents

C14. Density and Viscosity of Deuterium Oxide (Heavy Water)

C15. Densities and Specific Viscosities of Mixtures of Cesium Chloride in H2O and D2O at 0.0°C

C16. Values of β° for Several Gradient-Forming Salts

C17. Integrals for Computing Sedimentation Times through Linear Sucrose Gradients

Appendix D: Gradient Shapes

D1. Parameters for Isokinetic Gradients of Sucrose for Two IEC Rotors

D2. Parameters for Isokinetic Gradients in Sucrose for Four Spinco Rotors

D3. Equivolumetric Gradients of Sucrose for Oak Ridge B-XIV Rotors

D4. Equivolumetric Gradients of Sucrose for Oak Ridge B-XV Rotors

D5. Construction of Hyperbolic Gradient for the Separation of Ribosomes

D6. Acceleration Gradient

D7. Shapes of Sucrose and NaCI Gradients at Sedimentation-Diffusion Equilibrium

Appendix E: Zonal Rotors

E1. ORNL Designation for Zonal Rotors

Appendix F: Chemical Resistance of Various Plastics

Appendix G: Addresses of Some Manufacturers

Index

Details

No. of pages:
446
Language:
English
Copyright:
© Academic Press 1982
Published:
Imprint:
Academic Press
eBook ISBN:
9781483282060

About the Author

C. A. Price