Programmed Cell Death Methods and Protocols

This volume contains protocols specifically designed for studying programmed cell death, and also discusses recent advances in techniques that span broader areas of biology that have been recently used or that have potential to be incorporated into cell death research. The protocols are mostly described in the context of mammalian systems, but also cover other systems such as plants, Drosophila, and yeast. Programmed Cell Death: Methods and Protocols is comprised of 20 chapters: Chapters 1-5 describe apoptosis detection techniques; Chapter 6-9 describe methods for studying apoptosis associated with various pathologies in different organs including the lymphoid compartment, intestinal epithelium, granulocytes, and cardiomyocytes; Chapter 11-13 cover protocols and techniques for studying apoptosis in non-mammalian systems; Chapters 14-16 cover biochemical and biophysical methods for studying Bcl-2 family protein dynamics and protein-protein interactions during apoptosis;and the last four chapters explore protocols that are useful not only in apoptosis research but in wider areas of biological research, such as genome editing, inducible transgenes, and proteomics. Written in the highly successful Methods in Molecular Biology aeries format, chapters include introductions to their respective topics, lists of the necessary material and reagents, step-by-step, readily reproducible laboratory protocol, and tips on troubleshooting and avoiding known pitfalls.

Thorough and cutting-edge, Programmed Cell Death: Methods and Protocols is a comprehensive and valuable resource for researchers, ranging from beginner to expert, in their studies on programmed cell death.

Detection of Apoptotic Versus Autophagic Cell Death by Flow Cytometry.- In Vivo Apoptosis Imaging Using Site-Specifically ⁶⁸Ga-Labeled Annexin V.- Detection of Active Caspases during Apoptosis Using Fluorescent Activity-Based Probes.- Detection of Initiator Caspase Induced Proximity in Single Cells by Caspase Bimolecular Fluorescence Complementation.- In Vitro Use of Peptide Based Substrates and Inhibitors of Apoptotic Caspases.- Experimental In Vivo Sepsis Models to Monitor Immune Cell Apoptosis and Survival in Laboratory Mice.- Analysis of Cell Death Induction in Intestinal Organoids In Vitro.- In Vitro Differentiation of Mouse Granulocytes.- Hydrodynamic Injection as a Method of Gene Delivery in Mice: A Model of Chronic Hepatitis B Virus Infection.- Isolation of Cardiomyocytes and Cardiofibroblasts for Ex-Vivo Analysis.- Detection of Cell Death in Drosophila Tissues.- Methods to Study Plant Programmed Cell Death.- Modeling Metazoan Apoptotic Pathways in Yeast.- Characterizing Bcl-2 Family Protein Conformation and Oligomerization Using Cross-Linking and Antibody Gel-Shift in Conjunction with Native PAGE.- Using Förster-Resonance Energy Transfer to Measure Protein Interactions between Bcl-2 Family Proteins on Mitochondrial Membranes.- Preparing Samples for Crystallization of Bcl-2 Family Complexes.- Screening Strategies for TALEN Mediated Gene Disruption.- Using CRISPR/Cas9 Technology for Manipulating Cell Death Regulators.- Lentiviral Vectors to Analyze Cell Death Regulators.- Proteomic Profiling of Cell Death: Stable Isotope Labeling and Mass Spectrometry Analysis.