Centrifugation in Density Gradients

Centrifugation in Density Gradients

1st Edition - January 28, 1982

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  • Author: C. A. Price
  • eBook ISBN: 9781483282060

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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


    1 Particle Abstraction in Biology—An Introduction


    2 Origins of Density Gradient Centrifugation


    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


    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


    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


    6 Techniques of Preparative Density Gradient Centrifugation

    6.1 Planning a Separation

    6.2 Swinging-Bucket Separations

    6.3 Zonal Separations

    Data Processing


    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


    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


    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


    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


Product details

  • No. of pages: 446
  • Language: English
  • Copyright: © Academic Press 1982
  • Published: January 28, 1982
  • Imprint: Academic Press
  • eBook ISBN: 9781483282060

About the Author

C. A. Price

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