Apoptosis Methodsand Protocols

Hugh J . M. Brady

Preface v

Contributors i x

1 Measurement of Apoptosis by DNA Fragmentatio nDemetrius Matassov, Terri Kagan, Julie Leblanc,

Marianna Sikorska, and Zahra Zakeri 1

2 Measurement of Caspase Activity in Cells Undergoing Apoptosi sSharad Kumar 1 9

3 Flow Cytometry-Based Methods for Apoptosis Detectio nin Lymphoid Cell s

Owen Williams 31

4 Methods of Analyzing Chromatin Changes Accompanyin gApoptosis of Target Cells in Killer Cell Assay s

Richard C. Duke 43

5 Detecting and Quantifying Apoptosis in Tissue Section sVicki Save, Peter A. Hall, and Philip J. Coates 6 7

6 Immunoassay for Single-Stranded DNA in Apoptotic Cell sOskar S . Frankfurt 85

7 Methods to Measure Membrane Potential and Permeabilit yTransition in Mitochondria During Apoptosi s

Naoufal Zamzami and Guido Kroemer 103

8 Measurement of Changes in Intracellular Calciu mDuring Apoptosi s

David J. McConkey and Leta Nutt 11 7

9 Measurement of Changes in Apoptosis and Cell Cycl eRegulatory Kinase Cdk2

Gabriel Gil-Gömez 13 1

10 Assays to Measure Stress-Activated MAPK ActivityCarme Caelles and Monica Morales 145

11 Methods for Culturing Primary Sympathetic Neuron sand for Determining Neuronal Viabilit y

Jonathan Whitfield, Stephen J. Neame, and Jonathan Ham 157

12 Immunocytochemical Techniques for Studying Apoptosi sin Primary Sympathetic Neuron s

Stephen J. Neame, Jonathan Whitfield, and Jonathan Ham 169

13 Measuring Programmed Cell Death in Plant sLudmila Rizhsky, Vladimir Shulaev, and Ron Mittler 179

14 Detection of Apoptosis in DrosophilaKimberly McCall and Jeanne S. Peterson 191

15 Measurement of Apoptotic Cell Clearance In Vitr oAndrew Devitt and Christopher D . Gregory 207

16 Yeast Two-Hybrid Screening for Proteins InteractingWith the Anti-Apoptotic Protein A20

Karen Heyninck, Sofie Van Huffel, Marja Kreike,and Rudi Beyaert 223

17 The Yeast Three-Hybrid System As a Tool to Study Caspase sWim van Criekinge, Peter Schotte, Karen Heyninck,

and Rudi Beyaert 24318 Cloning of Apoptosis-Related Genes by Representationa l

Difference Analysis of cDN AMichael Hubank, Fredrik Bryntesson, Jennifer Regan,

and David G. Schatz 25519 Identification of Apoptosis Regulatory Genes Using

Insertional Mutagenesi sJoelle Thomas, Vann Leverrier, Anne-Laure Mathieu,

and Jacqueline Marvel 275Index 291