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Apoptosis Methods and Protocols

  • Book
  • © 2004

Overview

Editors:
  1. Hugh J. M. Brady

    1. Institute of Child Health, University College London, London, UK

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

Part of the book series: Methods in Molecular Biology (MIMB, volume 282)

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Table of contents (19 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Measurement of Apoptosis by DNA Fragmentation

    • Demetrius Matassov, Terri Kagan, Julie Leblanc, Marianna Sikorska, Zahra Zakeri
    Pages 1-17

  3. Measurement of Caspase Activity in Cells Undergoing Apoptosis

    • Sharad Kumar
    Pages 19-30

  4. Flow Cytometry-Based Methods for Apoptosis Detection in Lymphoid Cells

    • Owen Williams
    Pages 31-42

  5. Methods of Analyzing Chromatin Changes Accompanying Apoptosis of Target Cells in Killer Cell Assays

    • Richard C. Duke
    Pages 43-66

  6. Detecting and Quantifying Apoptosis in Tissue Sections

    • Vicki Save, Peter A. Hall, Philip J. Coates
    Pages 67-84

  7. Immunoassay for Single-Stranded DNA in Apoptotic Cells

    • Oskar S. Frankfurt
    Pages 85-101

  8. Methods to Measure Membrane Potential and Permeability Transition in the Mitochondria During Apoptosis

    • Naoufal Zamzami, Guido Kroemer
    Pages 103-115

  9. Measurement of Changes in Intracellular Calcium During Apoptosis

    • David J. McConkey, Leta Nutt
    Pages 117-130

  10. Measurement of Changes in Apoptosis and Cell Cycle Regulatory Kinase Cdk2

    • Gabriel Gil-Gómez
    Pages 131-144

  11. Assays to Measure Stress-Activated MAPK Activity

    • Carme Caelles, Mónica Morales
    Pages 145-156

  12. Methods for Culturing Primary Sympathetic Neurons and for Determining Neuronal Viability

    • Jonathan Whitfield, Stephen J. Neame, Jonathan Ham
    Pages 157-168

  13. Immunocytochemical Techniques for Studying Apoptosis in Primary Sympathetic Neurons

    • Stephen J. Neame, Jonathan Whitfield, Jonathan Ham
    Pages 169-178

  14. Measuring Programmed Cell Death in Plants

    • Ludmila Rizhsky, Vladimir Shulaev, Ron Mittler
    Pages 179-189

  15. Detection of Apoptosis in Drosophila

    • Kimberly McCall, Jeanne S. Peterson
    Pages 191-205

  16. Measurement of Apoptotic Cell Clearance In Vitro

    • Andrew Devitt, Christopher D. Gregory
    Pages 207-221

  17. Yeast Two-Hybrid Screening for Proteins Interacting With the Anti-Apoptotic Protein A20

    • Karen Heyninck, Sofie Van Huffel, Marja Kreike, Rudi Beyaert
    Pages 223-241

  18. The Yeast Three-Hybrid System As a Tool to Study Caspases

    • Wim van Criekinge, Peter Schotte, Karen Heyninck, Rudi Beyaert
    Pages 243-254

  19. Cloning of Apoptosis-Related Genes by Representational Difference Analysis of cDNA

    • Michael Hubank, Fredrik Bryntesson, Jennifer Regan, David G. Schatz
    Pages 255-273

  20. Identification of Apoptosis Regulatory Genes Using Insertional Mutagenesis

    • Joëlle Thomas, Yann Leverrier, Anne-Laure Mathieu, Jacqueline Marvel
    Pages 275-289
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About this book

The most fundamental question facing each and every cell within an org- ism is to survive or to die. Cell death is required for normal function; some estimates suggest that as many as one million cells undergo cell death every second in the adult human body. Almost all cells undergoing physiological, or programmed, cell death, independent of cell type, manifest a stereotypic p- tern of morphological changes termed apoptosis. Typically, apoptotic cells d- play shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation. The integrity of the cell membrane is not lost during apoptosis and so avoids eliciting the inflammatory response that would have been caused by the spillage of the cell’s contents. This is quite in contrast to the loss of cell contents typical of necrosis. The caspases, the family of intracellular cysteine proteases associated with apoptosis, are responsible for the stereotypical m- phological changes. Caspases cleave various substrate proteins that act on DNA fragmentation, nuclear envelope integrity, the cytoskeleton, and cell volume regulation. Apoptotic cells are cleared in vivo by the process of phagocytosis, in which specific “phagocytes” move to the site of apoptosis, engulf the dying cells and digest them. Apoptosis has a central role in many physiological processes, for example, in the immune system. Autoreactive cells are deleted via apoptosis to prevent autoimmunity. At the end of an immune response, activated lymphocytes are removed to maintain homeostasis within the immune system.

Reviews

"The calibre of the contributors to this book, make it an authoritative tome. As it also provides insights in to how and why specific protocols should be applied, it will be a great asset to those studying apoptosis in laboratory-based haematology." -British Journal of Haematology

"...well documented...clearly laid out...a daily reference to the laboratory for post graduate students." - Veterinary Pathology

Editors and Affiliations

  • Institute of Child Health, University College London, London, UK

    Hugh J. M. Brady

Bibliographic Information

  • Book Title: Apoptosis Methods and Protocols

  • Editors: Hugh J. M. Brady

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1385/1592598129

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2004

  • eBook ISBN: 978-1-59259-812-0Published: 05 February 2008

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 304

  • Number of Illustrations: 74 b/w illustrations

  • Topics: Cell Biology

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