The ability to detect and quantify apoptosis, to understand its biochemistry, and to identify its regulatory genes and proteins is crucial to biomedical research. In Apoptosis: Methods and Protocols, Second Edition, expert researchers describe the techniques to best investigate the critical steps involved in the apoptotic process. Presented from several different research perspectives, the volume contains sections covering detection of apoptosis, detection of non-apoptotic cell death, modifications of apoptotic proteins during apoptosis, the analysis of its major regulators, as well as analysis of apoptosis in different organs and in model organisms. Written in the highly successful Methods in Molecular Biology'?? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.Comprehensive and cutting-edge, Apoptosis: Methods and Protocols, Second Edition constitutes a key technical reference to the significant methodologies used in the field, and offers beginners and experienced researchers powerful tools to illuminate the phenomena of programmed cell death.

Preface p. v
Contributors p. ix
Detection of Apoptosis
Analysing Caspase Activation and Caspase Activity in Apoptotic Cells p. 3
Flow Cytometry-Based Apoptosis Detection p. 19
Live to Dead Cell Imaging p. 33
Detection of Apoptosis in Tissue Sections p. 49
Detection of Apoptosis in Cell-Free Systems p. 65
Detection of Non-Apoptotic Cell Death
Methods to Analyze Cellular Necroptosis p. 79
Detection of Cell Death by Autophagy p. 95
Modifications of Apoptotic Proteins During Apoptosis
Methods to Analyze Transglutamination of Proteins Involved in Apoptosis p. 107
Methods to Analyze S-nitrosylation of Proteins Involved in Apoptosis p. 117
Application of In Vivo EPR for Tissue pO2 and Redox Measurements p. 131
Analysis of the Function of Major Regulators of Apoptosis/Cell Survival
Assays to Measure p53-Dependent and -Independent Apoptosis p. 143
Measurement of Changes in Cdk2 and Cyclin O-Associated Kinase Activity in Apoptosis p. 161
Fluorometric Methods for Detection of Mitochondrial Membrane Permeabilization in Apoptosis p. 173
Regulation of Apoptosis by the Unfolded Protein Response p. 191
Detection of Uncoupling Protein-2 (UCP2) As a Mitochondrial Modulator of Apoptosis p. 205
Multiple Approach to Analyzing the Role of MicroRNAs in Apoptosis p. 219
Assessment of Apoptotic Cell Phagocytosis by Macrophages p. 247
Analysis of Apoptosis in Different Organs
Detection of Apoptosis in Mammalian Development p. 259
Detection of Apoptosis in the Central Nervous System p. 273
Genetic Mapping of Anti-Apoptosis Pathways in Myeloid Progenitor Cells p. 283
Analysis of Apoptosis in Isolated Primary Cardiac Myocytes p. 293
Cell Death in Myoblasts and Muscles p. 313
Analysis of Apoptosis in Model Organisms
Reliable Method for Detection of Programmed Cell Death in Yeast p. 335
Detection of Cell Death in Drosophila p. 343
Detecting Apoptotic Cells and Monitoring Their Clearance in the Nematode Caenorhabditis elegans p. 357
Detection of Herpes Simplex Virus Dependent Apoptosis p. 371
Index p. 389
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