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This four-volume laboratory manual contains comprehensive state-of-the-art protocols essential for research in the life sciences. Techniques are presented in a friendly step-by-step fashion, providing useful tips and potential pitfalls. The important steps and results are beautifully illustrated for further ease of use. This collection enables researchers at all stages of their careers to embark on basic biological problems using a variety of technologies and model systems. This thoroughly updated third edition contains 165 new articles in classical as well as rapidly emerging technologies.
Topics covered include:
- Cell and Tissue Culture: Associated Techniques, Viruses, Antibodies, Immunocytochemistry (Volume 1)
- Organelle and Cellular Structures, Assays (Volume 2)
- Imaging Techniques, Electron Microscopy, Scanning Probe and Scanning Electron Microscopy, Microdissection, Tissue Arrays, Cytogenetics and In Situ Hybridization, Genomics and Transgenic Knockouts and Knock-down Methods (Volume 3)
- Transfer of Macromolecules, Expression Systems, Gene Expression Profiling (Volume 4)
- Indispensable bench companion for every life science laboratory
- Provides the latest information on the plethora of technologies needed to tackle complex biological problems
- Includes numerous illustrations, some in full color, supporting steps and results
Cell biologists, molecular biologists, biochemists, biotechnologists, geneticists, neuroscientists, microbiologists, developmental biologists, toxicologists, and virologists
Contents of Volume 1
Part A. Cell and Tissue Culture: Associated Techniques.
Section 1. General Techniques.
Section 2. Culture of Specific Cell Types: Stem Cells.
Section 3. Culture of Specific Cell Types: Haemopoietic, Mesenchymal, and Epithelial.
Section 4. Differentiation and Reprogramming of Somatic Cells.
Section 5. Immortalisation.
Section 6. Somatic Cell Hybrids.
Section 7. Cell Separation Techniques.
Section 8. Cell Cycle Analysis.
Section 9. Cytotoxic and Cell Growth Assays.
Section 10. Apoptosis.
Section 11. Assays of Cell Transformation, Tumorigenesis, Invasion and Wound Healing.
Section 12. Electrophysiological Methods.
Section 13. Organ Cultures.
Part B. Viruses.
Section 14. Growth and Purification of Viruses.
Part C. Antibodies.
Section 15. Production and Purification of Antibodies.
Part D. Immunocytochemistry.
Section 16. Immunofluoresence.
Part E. Appendix.
Contents of Volume 2
Part A. Organelles and Cellular Structures.
Section 1. Isolation: Plasma Membrane, Organelles, and Cellular Structures.
Section 2. Vital Staining of Cells/Organelles.
Section 3. Protein Purification.
Part B. Assays.
Section 4. Endocytic and Exocytic Pathways.
Section 5. Membranes.
Section 6. Mitochondria.
Section 7. Nuclear Transport.
Section 8. Chromatin Assembly.
Section 9. Signal Transduction Assays.
Section 10. Assays and Models of in Vitro and in Vitro Motility.
Section 11. Mechanical Stress in Single Cells.
Part C. Appendix.
Contents of Volume 3
Part A. Imaging Techniques.
Section 1. Light Microscopy.
Section 2. Digital Video Microscopy.
Section 3. Confocal Microscopy of Living Cells and Fixed Cells.
Section 4. Fluorescent Microscopy of Living Cells.
Section 5. Use of Fluorescent Dyes for Studies of Intracellular Physiological Parameters.
Section 6. Digital Image Processing, Analysis, Storage, and Display.
Part B. Electron Microscopy.
Section 7. Specimen Preparation Techniques.
Section 8. Cryotechniques.
Section 9. Electron Microscopy Studies of the Cytoskeleton.
Section 10. Immunoelectron Microscopy.
Part C. Scanning Probe and Scanning Electron Microscopy.
Section 11. Scanning Probe and Scanning Electron Microscopy.
Part D. Microdissection.
Section 12. Tissue and Chromosome Microdissection.
Part E. Tissue Arrays.
Section 13. Tissue Arrays.
Part F. Cytogenetics and in Situ Hybridisation
Section 14. Cytogenetics.
Section 15. In Situ Hybridisation.
Part G. Genomics.
Section 16. Genomics.
Part H. Transgenic, Knockouts, and Knockdown Methods.
Section 17. Transgenic, Knockouts and knock-down methods.
Contents of Volume 4
Part A. Transfer of Macromolecules.
Section 1. Proteins.
Section 2. Genes.
Section 3. Somatic Cell Nuclear Transfer.
Part B. Expression Systems.
Section 4. Expression Systems.
Part C. Gene Expression Profiling.
Section 5. Differential Gene Expression.
Part D. Proteins.
Section 6. Protein Determination and Analysis.
Section 7. Sample Fractionation for Proteomics.
Section 8. gel Electrophoresis.
Section 9. Detection of Proteins in Gels.
Section 10. Gel Profiling of Posttranslationally Modified Proteins.
Section 11. Protein/Protein and Protein/Small Molecule Interactions.
Section 12. Functional Proteomics.
Section 13. Protein/DNA Interactions.
Section 14. Protein Degradation.
Section 15. Mass Spectrometry: Protein Identification and Interactions.
Part E. Appendix.
- No. of pages:
- © Academic Press 2006
- 9th November 2005
- Academic Press
- Hardcover ISBN:
- eBook ISBN:
A native of Chile, Dr. Julio E. Celis received his bachelor’s degree from the University of Chile in 1964, and then completed his Ph.D. in the United States at the University of Iowa. After his postdoctoral training at the Medical Research Council Laboratory of Molecular Biology in Cambridge, England, he went on to become an assistant professor at the University of Chile. Dr. Celis was named an associate professor in 1975 at Aarhus University, Denmark, and in 1987 became Chairman of the University’s Institute of Medical Biochemistry. He is also the Chairman of the Danish Centre for Human Genome Research and was recently elected Secretary General-elect of the Federation of European Biochemical Societies (FEBS) 1997. With nearly 180 publications to his credit, Dr. Celis’ specialized areas of research include molecular mechanisms of cancer, human 2D gel protein databases and their link to genome data, signal transduction, and the biology of human skin.
Married, with three children and one cat (Max William), Dr. Celis currently resides in Denmark where he holds among his many titles the chair to the Symposium and Prize Committee of the Danish Cancer Society, as well as the acting Chairman of the Nordic Molecular Biology Association (NOMBA). Dr. Celis is currently the Vice President of the European Molecular Biology Laboratory (EMBL) Council and leader of the Danish Delegation to the OECD Megascience Forum subgroup on Bioinformatics. He is also the European Union Observer to the International Nucleotide sequence database collaboration and a member of the EMBO Course Committee.
Danish Cancer Society; Institute of Cancer Biology and Danish Centre for Human Genome Research; Copenhagen, Denmark
Nigel Carter is the Head of Molecular Cytogenetics at the Sanger Centre, Cambridge UK and is currently the Secretary of the International Society for Analytical Cytology. Receiving his BA and D.Phil degrees from the University of York where he specialised in parasitology, Nigel became interested in flow cytometry when he was appointed to the Nuffield Department of Surgery at the University of Oxford in 1981. In 1989, Nigel took up a post in the Department of Pathology, University of Cambridge to use molecular cytogenetic technologies to study human karyotype abnormalities. Nigel's work in this field has involved the application of digital microscopy to fluorescence in situ hybridisation and the development of flow cytometry for chromosome sorting for the generation of chromosome-specific DNA libraries and paints.
The Sanger Center, Welcome Trust, Cambridge, United Kingdom
Max-Planck Institute of Molecular and Cell Biology and Genetics, Munich, Germany
J. Victor Small received a Ph.D. in 1969 at Kings College, London from where he moved to Denmark to take up a lectureship at Aarhus University, which he held until 1977. Thereafter, he became department head in a new institute, the Institute of Molecular Biology, established by the Austrian Academy of Sciences in Salzburg, of which he is the current director. His work has centered on smooth muscle biochemistry and structure and on the mechanism of cell motility.
Austria Academy of Sciences, Salzburg, Austria
Tony Hunter received his Ph.D. in 1969 from the University of Cambridge, England. He joined the Salk Institute in 1975 as an assistant professor and has been a professor since 1982. His current interests are the protein-tyrosine kinases of the Src and growth factor receptor families and the protein-tyrosine phosphatases that remove the phosphates added by protein-tyrosine kinases. He was elected as a Fellow of the Royal Society of London in 1987, a Fellow of the American Academy of Arts and Sciences in 1992, and as an Associate Member of the European Molecular Biology Organization in 1992.
The Salk Institute, La Jolla, California, USA
David Shotton, since 1981 a University Lecturer in Cell Biology at the University of Oxford, graduated in biochemistry from Cambridge University in 1965. During his doctoral research at the MRC Laboratory of Molecular Biology, Cambridge (1965-1969), he completed the sequence and crystallographic structure determination of the enzyme elastase. Following postdoctoral studies at the University of Bristol, Berkeley, and Harvard, during which he changed fields to cell biology, studying membrane structure, he took a lectureship at Imperial College in 1976. His current interests include cellular immunology, advanced light and electron microscopy techniques, digital image processing, and hypermedia.
University of Oxford, U.K.
"4 Stars - These 4 volumes provide a modern, near encyclopedic review of cell biology protocols." --DOODY'S, May 2006
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