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Density Gradient Centrifugation
- 1st Edition, Volume 6 - January 1, 1978
- Editors: T. S. Work, E. Work
- Language: English
- eBook ISBN:9 7 8 - 0 - 0 8 - 0 8 5 8 7 5 - 3
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Part I R. Hinton and M. Dobrota Density Gradient Centrifugation
Density Gradient Centrifugation
Chapter 1: Introduction to zonal centrifugation
1.1. The first applications of centrifugation in biology
1.2. Centrifugal techniques
1.3. The development of centrifuges and rotors
1.4. Uses and limitations of centrifugal techniques
1.5. Design of a centrifuge laboratory
1.6. Safety of centrifuges
1.7. Care of rotors
1.8. Guarantees on rotors
Chapter 2: Theoretical aspects of centrifugal separations
2.1. Theory of rate-zonal separations
2.2. Theory of isopycnic banding
2.3. Effects of density gradient solutes on subcellular structures
Chapter 3: Conditions for a centrifugal separation
3.1. Choice of approach
3.2. Choice of rotor
3.3. Density gradient solutes
3.4. Choice of gradient
Chapter 4: Centrifugation in conventional rotors
4.1. Rate-zonal centrifugation
4.2. Isopycnic zonal centrifugation
Chapter 5: Centrifugation in zonal rotors
5.2 Conventional, non-reorienting zonal rotors
5.2 Reorienting zonal rotors
Chapter 6: Assay of fractions separated by density gradient centrifugation
6.1. Enzyme and chemical assays on fractions
6.2. Electron microscopic examination of fractions
6.3. Assessment of results from density gradient separations
6.4. Calculation of sedimentation coefficients
Chapter 7: Applications of density gradient centrifugation
7.1. Separation of living cells
7.2. Separation of cell organelles from mammalian tissues
7.3. Separation of subcellular structures from plant cells
7.4. Separation of subcellular components from unicellular organisms
7.5. Fractionation of macromolecules
7.6. Other applications of density gradient centrifugation in biochemistry
7.7. Other applications of density gradient centrifugation
Chapter 8: Artefacts arising during centrifugal separations
8.1. Damage to particles during centrifugation
8.2. Factors affecting the accuracy of assays performed on fractions from density gradients
8.3. Uncertainties in estimates of particle density and sedimentation coefficient
Chapter 9: Future prospects for density gradient centrifugation
9.1. Centrifuge design
9.2. Developments in centrifuge rotors
9.3. Developments in ancillary systems
9.4. Uses of centrifugal methods
Note added in proof
Acknowledgements
Appendices
Appendix I
Appendix II
Appendix III
Appendix IV
References
Subject index
Part II T. Chard An Introduction to Radioimmunoassay and Related Techniques
An Introduction to Radioimmunoassay and Related Techniques
List of abbreviations
Chapter 1: The background to radioimmunoassay
1.1. Introduction
1.2. Terminology
1.3. Early development of radioimmunoassay
1.4. Basic principles of binding assays
1.5. Binder dilution curves and standard curves
1.6. Methods of plotting the standard curve
1.7. The importance of K value
1.8. The measurement of K value
1.9. A model system for binding assays
1.10. Some implications of the model system
Chapter 2: Requirements for a binding assay – purified ligand
2.1. Requirements for a binding assay
2.2. The need for purified ligand
2.3. Availability of pure ligand
2.4. Dissimilarity between purified ligand and endogenous ligand
2.5. Standards
2.6. Storage of materials
Chapter 3: Requirements for binding assays – tracer ligand
3.1. Radioactive isotopes
3.2. Counting of radioactive isotopes
3.3. Choice of a counter
3.4. Some practical aspects of isotope counting
3.5. Essential characteristics of a tracer
3.6. Preparation of tracers
3.7. Iodinated tracers
3.8. Alternative labels for tracers
Chapter 4: Requirements for a binding assay – the binder
4.1. Characteristics required of a binder
4.2. Antibodies
4.3. Cell receptors
4.4. Circulating binding proteins
4.5. Radioassay for the detection of endogenous antibodies and circulating binding proteins
Chapter 5: Requirements for a binding assay – separation of bound and free ligand
5.1. Efficiency of separation methods
5.2. Practicality of separation methods
5.3. Methods for the separation of bound and free ligand
5.4. Immunoradiometric techniques
Chapter 6: Requirements for a binding assay – extraction of ligand from biological fluids
6.1. Extraction for concentration of ligand
6.2. Extraction for purification of ligand
6.3. General aspects of extraction procedures
Chapter 7: Requirements for binding assays – calculation of results
7.1. Calculation of results by simple manual extrapolation
7.2. Linearisation of the standard curve
7.3. Electronic aids to calculation of results
7.4. Estimation of confidence limits to the results
Chapter 8: Characteristics of binding assays – sensitivity
8.1. Definition of sensitivity
8.2. Methods of increasing the sensitivity of a binding assay
8.3. Methods of decreasing the sensitivity of an assay
8.4. Targeting of binding assay – the importance of ranges
8.5. Optimisation of an assay by theoretical analysis
8.6. Conclusions
Chapter 9: Characteristics of binding assays – specificity
9.1. Definition of specificity
9.2. Specific non-specificity
9.3. Non-specific non-specificity
Chapter 10: Characteristics of binding assays – precision
10.1. Definitions
10.2. Factors affecting precision
10.3. Methods for monitoring the precision of a binding assay
10.4. Methods for optimising the precision of a binding assay
Chapter 11: Characteristics of binding assays – relation to other types of assay
11.1. Definition
11.2. Receptor assays
11.3. Assays using circulating binding proteins
11.4. Immunoassays
11.5. Conclusions
Chapter 12: Automation of binding assays
12.1. General
12.2. Identification and dispensing of the sample
12.3. Addition of reagents
12.4. Incubation
12.5. Separation of bound and free ligand
12.6. Counting of radioactivity
12.7. Calculation of results
12.8. Conclusions
Chapter 13: Organisation of assay services
13.1. Who should perform radioimmunoassay?
13.2. Organisation of an assay laboratory
13.3. Organisation of assay services
Appendices
Appendix I
Appendix II
Appendix III
Appendix IV
Appendix V
References
Subject index
- No. of pages: 533
- Language: English
- Edition: 1
- Volume: 6
- Published: January 1, 1978
- Imprint: North Holland
- eBook ISBN: 9780080858753
TW
T. S. Work
Affiliations and expertise
N.I.M.R., Mill HillEW
E. Work
Affiliations and expertise
Imperial College, LondonRead Density Gradient Centrifugation on ScienceDirect