Apoptosis (eBook)

Front Cover 1
Apoptosis 4
Copyright Page 5
Table of Contents 6
Contributors to Volume 322 10
Preface 14
Volumes in Series 16
Section I: Measuring Apoptosis and Apoptosis-Induced Endonucleases 35
Chapter 1. Detection of DNA Cleavage in Apoptotic Cells 37
Chapter 2.Detection of Apoptosis by Annexin V Labeling 49
Chapter 3. Analysis of Apoptotic Cells by Flow and Laser Scanning Cytometry 52
Chapter 4. Quantitative Measurement of Apoptosis Induced by Cytotoxic T Lymphocytes 74
Chapter 5. Apoptotic Nuclease Assays 81
Section II: Measuring Apoptosis in Lower Organisms 97
Chapter 6. Analysis of Programmed Cell Death and Apoptosis in Drosophila 99
Chapter 7. Programmed Cell Death in the Nematode Caenorhabditis elegans 110
Section III: Proteases Involved in Apoptosis and Their Inhibitors 123
Chapter 8. Caspase Assays 125
Chapter 9. Determination of Caspase Specificities Using a Peptide Combinatorial Library 134
Chapter 10. Criteria for Identifying Authentic Caspase Substrates during Apoptosis 144
Chapter 11. Purification and Use of Granzyme B 159
Chapter 12. Viral Caspase Inhibitors CrmA and p35 177
Chapter 13. Purification and Use of Recombinant Inhibitor of Apoptosis Proteins as Caspase Inhibitors 188
Chapter 14. Monitoring Activity of Caspases and Their Regulators in Yeast Saccharomyces cerevisiae 196
Section IV: Cell-Free Systems for Monitoring Steps in Apoptosis Pathways 209
Chapter 15. In Vitro Assays for Caspase-3 Activation and DNA Fragmentation 211
Chapter 16. Cell-Free Apoptosis in Xenopus laevis Egg Extracts 217
Chapter 17. Cytofluorometric Quantification of Nuclear Apoptosis Induced in a Cell-Free System 232
Section V: Mitochondria and Apoptosis 237
Chapter 18. Purification of Mitochondria for Apoptosis Assays 287
Chapter 19. Quantitation of Mitochondrial Transmembrane Potential in Cells and in Isolated Mitochondria 241
Chapter 20. Nitrogen Cavitation for Cell Disruption to Obtain Mitochondria from Cultured Cells 246
Chapter 21. Apoptosis-Related Activities Measured with Isolated Mitochondria and Digitonin-Permeabilized Cells 255
Chapter 22. Assays for Cytochrome c Release from Mitochondria during Apoptosis 268
Chapter 23. Purification and Liposomal Reconstitution of Permeability Transition Pore Complex 276
Section VI: Bcl-2 Family Proteins 286
Chapter 24. Monitoring Interactions of Bcl-2 Family Proteins in 96-Well Plate Assays 289
Chapter 25. Analysis of Dimerization of Bcl-2 Family Proteins by Surface Plasmon Resonance 300
Chapter 26. Measuring Pore Formation by Bcl-2 Family Proteins 308
Chapter 27. Assays for Studying Bax-Induced Lethality in the Yeast Saccharomyces cerevisiae 317
Chapter 28. Exploiting the Utility of Yeast in the Context Programmed Cell Death 331
Section VII: Studying Receptors and Signal Transduction Events Implicated in Cell Survival and Cell Death 357
Chapter 29. Production of Recombinant TRAIL and TRAIL Receptor: Fc Chimeric Proteins 359
Chapter 30. Expression of Lymphotoxins and Their Receptor–Fc Fusion Proteins by Baculovirus 379
Chapter 31. Analysis of the CD95 (APO-1/Fas) Death-Inducing Signaling Complex by High-Resolution Two-Dimensional Gel Electrophoresis 397
Chapter 32. Measurement of Ceramide Levels by the Diacylglycerol Kinase Reaction and by High-Performance Liquid Chromatography–Fluorescence Spectrometry 407
Chapter 33. Measurement of Ceramide Synthase Activity 412
Chapter 34. Measurement of Sphingomyelinase Activity 416
Chapter 35. Assays for JNK and p38 Mitogen-Activated Protein Kinases 422
Chapter 36. Assaying for IKB Kinase Activity 427
Chapter 37. Assays for Akt 434
Section VIII: Other Methods 445
Chapter 38. Measurement of Cellular Oxidation, Reactive Oxygen Species, and Antioxidant Enzymes during Apoptosis 447
Chapter 39. Volume Regulation and Ion Transport during Apoptosis 455
Chapter 40. Assays for Transglutaminases in Cell Death 467
Chapter 41. Anoikis 506
Chapter 42. Transient Transfection Assay of Cell Death Genes 514
Chapter 43. Sindbis Virus Vector System for Functional Analysis of Apoptosis Regulators 526
Chapter 44. Transduction of Full-Length Tat Fusion Proteins Directly into Mammalian Cells: Analysis of T Cell Receptor Activation-Induced Cell Death 542
Author Index 557
Subject Index 589