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Centrifugal Separations in Molecular and Cell Biology
1st Edition - July 13, 1978
Editors: G.D. Birnie, D. Rickwood
Language: English
eBook ISBN:9781483278414
9 7 8 - 1 - 4 8 3 2 - 7 8 4 1 - 4
Centrifugal Separations in Molecular and Cell Biology focuses on the application of modern centrifugation technology in molecular and cell biology, including the separation and…Read more
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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.
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