Centrifugal Separations in Molecular and Cell Biology

Centrifugal Separations in Molecular and Cell Biology

1st Edition - July 13, 1978
This is the Latest Edition
  • Editors: G.D. Birnie, D. Rickwood
  • eBook ISBN: 9781483278414

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Description

Centrifugal Separations in Molecular and Cell Biology focuses on the application of modern centrifugation technology in molecular and cell biology, including the separation and fractionation of biological particles by centrifugation on the preparative and analytical scales. The selection first covers the principles and practices of centrifugation and the bases of centrifugal separations. Discussions focus on the basic concepts of sedimentation theory, centrifugation methods, designing centrifugation experiments, care of centrifuges and rotors, and statistical estimation of molecular parameters. The book also ponders on the practical aspects of rate-zonal centrifugation, including gradient materials, density and viscosity of glycerol solutions, and resolution and gradient shape. The publication examines fractionations in zonal rotors and the quantitative aspects of rate-zonal centrifugation. The text then reviews isopycnic centrifugation in ionic media and analytical centrifugation. Topics include separation by isopycnic banding, large-scale preparative procedures, and density-gradient solutes. The selection is a valuable reference for readers interested in centrifugation technology.

Table of Contents


  • Contents

    1 Ιntroduction: Principles and Practices of Certification

    Basic Concepts of Sedimentation Theory

    Centrifugation Methods

    Designing Centrifugation Experiments

    Care of Centrifuges and Rotors

    2 The Bases of Centrifugal Separations

    Comparison of Zonal Centrifugation With Complementary Techniques

    Differential Versus Zonal Centrifugation

    Theory of Rate-Zonal Experiments

    Sedimentation Coefficient

    Location of the Centroid

    Resolution in Velocity Experiments

    Isopycnic Zonal Experiments

    Time to Reach Equilibrium

    Resolution of Isopycnic Zones

    Interaction Between Solutes in Isopycnic Experiments

    Statistical Estimation of Molecular Parameters

    S-Values

    Statistical Procedures

    Simulation of Zonal Experiments

    Appendix

    Relationships Between s and Radial Co-Ordinates

    Convective Instabilities in Zones

    Isopycnic Equilibrium

    Least-Squares Analysis For Curvilinear Polynomials

    References

    3 Practical Aspects of Rate-Zonal Centrifugation

    Experimental Design

    Gradient Materials

    Simple Sugars

    Glycerol

    Polysaccharides

    Proteins

    Deuterium Oxide

    Mixed-Solute Gradients

    Resolution and Gradient Shape

    Resolution

    Design of Gradients

    Gradient Generation

    Loading, Running and Analyzing Gradients

    Sample Loading

    Centrifugation Conditions

    Unloading and Analysis

    Appendix I: Density and Viscosity of Sucrose Solutions

    Appendix Ii: Density and Viscosity of Glycerol Solutions

    References

    4 Fractionations in Zonal Rotors

    Design and Operation of Batch-Type Rotors

    Dynamic Loading and Unloading

    Statically-Loaded (Unloaded) Rotors

    Gradients and Sample Preparation

    Fractionations in Batch-Type Rotors

    Separations of Different Cell Types

    Separation of Cells According to Position in The Cell Cycle

    Nuclei and Nuclear Membranes

    Mitochondria, Lysosomes and Peroxisomes

    Plasma Membrane and Endoplasmic Reticulum

    Ribosomes and Polysomes

    Nucleic Acids

    Cell Surface Coat (Glycocalyx)

    Plasma Lipoproteins

    Viruses

    Continuous-Flow Zonal Centrifugation

    Design and Operation of Rotors

    Fractionation With B-Type Rotors

    Fractionation With J- and K-Series Rotors

    Centrifugal Elutriation

    References 1

    5 Rate-Zonal Centrifugation: Quantitative Aspects

    Some Important Underlying Assumptions

    Guide For The Computerized Calculation of Sedimentation Coefficients

    Guide For The Manual Calculation of Sedimentation Coefficients

    Accuracy of Sedimentation Coefficients Calculated From Density-Gradient Data Using Large-Scale Zonal Rotors

    Some Important Sources of Error

    Other Analytical Uses

    Appendix I: FORTRAN Program For Calculating Sedimentation Coefficients

    Appendix Ii: Calculating Radius-Volume Relationships

    Appendix Iii: Sedim Values For Various Particle Densities

    Appendix Iv: Calculating The Natural Logarithm of the Rotor Radius

    References

    6 Isopycnic Centrifugation in Ionic Media

    Basic Procedures

    Optimum Design of Buoyant Density-Gradient Separations

    Equilibrium Gradients

    Non-Equilibrium Preformed Gradients

    Resolving Power of Gradients

    Fractionation of Gradients

    Buoyant Density-Gradient Solutes

    Data From Isopycnic Gradients

    Separations By Isopycnic Banding

    Large-Scale Preparative Procedures

    Isolation of DNA

    Isolation of RNA

    Isolation of Protein

    References

    7 Isopycnic Centrifugation in Non-Ionic Media

    General Techniques

    Density-Gradient Solutes

    Monosaccharides and Disaccharides

    Polysaccharides

    Lodinated Compounds

    Colloidal Silica

    Other Gradient Media

    References

    8 Analytical Ultracentrifugation

    History and Scope

    Instrumentation

    Optical Systems

    Analytical Ultracentrifuge Cells

    Sedimentation Equilibrium Analysis

    Sedimentation Equilibrium in Two-Component Systems

    Sedimentation Equilibrium Analysis of Associating Systems

    Sedimentation Equilibrium in Multi-Component Systems

    Sedimentation Velocity Analysis

    Areas of Application

    Measurement of Sedimentation Coefficient

    Use of Sedimentation Coefficients in Molecular-Weight Determination

    Sedimentation Velocity Analysis and Molecular Shape

    Large Molecule-Small Molecule Interactions

    Large Molecule-Large Molecule Interactions

    Conclusions

    References

    9 Characteristics of Ultracentrifuge Rotors and Tubes

    Designs and Types of Rotor

    Materials

    Types of Rotor

    Stability of Rotors

    Centrifuge Tubes and Caps

    Tubes

    Sealing Caps For Tubes

    Derating Rotors

    Routine Maintenance of Rotors

    Appendix: Physical Dimensions and Characteristics of Rotors

    The Relationship Between Radius and Volume of Zonal Rotors

    Index


Product details

  • No. of pages: 340
  • Language: English
  • Copyright: © Butterworth-Heinemann 1978
  • Published: July 13, 1978
  • Imprint: Butterworth-Heinemann
  • eBook ISBN: 9781483278414

About the Editors

G.D. Birnie

D. Rickwood

Affiliations and Expertise

University of Essex, Wivenhoe Park, Colchester