ABSTRACT
Protocol 11.3: Pl staining of nuclei in cell monolayers 301
Protocol 11.4: Hoechst 33342/PI staining of cells in culture 302
Protocol 11.5: AO staining to assay apoptosis in Drosophila embryos
11.4 Proteolysis of nuclear proteins 303
Protocol 11.6: Western blotting analysis of PARP cleavage 304
11.5 DNA fragmentation in apoptosis 306
Protocol 11.7: Localisation of EndoG or AIF in cell nuclei 307
Protocol 11.8: Detection of nucleosome ladders by agarose gel electrophoresis
Protocol 11.9: Detection of high molecular weight DNA fragments by gel electrophoresis
Protocol 11.10: Detection of low molecular weight DNA laddering in cultured cells
Protocol 11.11: TUNEL analysis of fragmented DNA in whole mount Drosophila embryos
Protocol 11.12: In situ end-labelling of fragmented DNA in cell monolayers
Protocol 11.13: In situ end-labelling of tissue sections 314
Protocol 11.14: Comet assay for detection of apoptotic and necrotic nuclei
11.6 Concluding remarks 320
11.7 References 321
11.1 Introduction
The role of apoptosis in the programmed cell death that is essential for normal embryonic development is now well established [1]. Moreover, an expanding number of clinical disorders have been identified in which apoptosis forms a major component [2]. These include neurodegenerative disorders such as Alzheimer’s disease and ischaemic stroke, autoimmune diseases and cancer. Apoptosis involves a regulated series of events to efficiently terminate the cellular function and to dismantle the cell while maintaining homeostasis. To accomplish this complicated task, critical organelles and structures are rapidly modified.
