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This volume containsprotocols specifically designed for studying programmed cell death, and alsodiscusses recent advances in techniques that span broader areas of biology thathave been recently used or that have potential to be incorporated into cell deathresearch. The protocols are mostly described in the context of mammaliansystems, but also cover other systems such as plants, Drosophila, andyeast. Programmed Cell Death: Methods and Protocols is comprised of 20chapters: Chapters 1-5 describe apoptosis detection techniques; Chapter 6-9describe methods for studying apoptosis associated with various pathologies indifferent organs including the lymphoid compartment, intestinal epithelium,granulocytes, and cardiomyocytes; Chapter 11-13 cover protocols and techniquesfor studying apoptosis in non-mammalian systems; Chapters 14-16 coverbiochemical and biophysical methods for studying Bcl-2 family protein dynamicsand protein-protein interactions during apoptosis;and the last four chaptersexplore protocols that are useful not only in apoptosis research but in widerareas of biological research, such as genome editing, inducible transgenes, andproteomics. Written in the highly successful Methods in Molecular Biologyaeries format, chapters include introductions to their respective topics, listsof the necessary material and reagents, step-by-step, readily reproduciblelaboratory protocol, and tips on troubleshooting and avoiding known pitfalls.
Thoroughand cutting-edge, Programmed Cell Death: Methods and Protocols is acomprehensive and valuable resource for researchers, ranging from beginner toexpert, in their studies on programmed cell death.
List of contents
Detection of Apoptotic Versus Autophagic Cell Death by Flow Cytometry.- In Vivo Apoptosis Imaging UsingSite-Specifically 68Ga-Labeled Annexin V.- Detection of Active Caspasesduring Apoptosis Using Fluorescent Activity-Based Probes.- Detection of Initiator CaspaseInduced Proximity in Single Cells by Caspase Bimolecular Fluorescence Complementation.- In Vitro Use of Peptide Based Substratesand Inhibitors of Apoptotic Caspases.- Experimental In Vivo Sepsis Modelsto Monitor Immune Cell Apoptosis and Survival in Laboratory Mice.- Analysis of Cell Death Inductionin Intestinal Organoids In Vitro.- In Vitro Differentiation of MouseGranulocytes.- Hydrodynamic Injection as a Methodof Gene Delivery in Mice: A Model of Chronic Hepatitis B Virus Infection.- Isolation of Cardiomyocytesand Cardiofibroblasts for Ex-Vivo Analysis.- Detection of Cell Death in Drosophila Tissues.- Methods to Study Plant ProgrammedCell Death.- Modeling Metazoan Apoptotic Pathwaysin Yeast.- Characterizing Bcl-2 Family ProteinConformation and Oligomerization Using Cross-Linking and Antibody Gel-Shift in Conjunctionwith Native PAGE.- Using Förster-Resonance EnergyTransfer to Measure Protein Interactions between Bcl-2 Family Proteins on MitochondrialMembranes.- Preparing Samples for Crystallizationof Bcl-2 Family Complexes.- Screening Strategies forTALEN Mediated Gene Disruption.- Using CRISPR/Cas9 Technologyfor Manipulating Cell Death Regulators.- Lentiviral Vectors to AnalyzeCell Death Regulators.- Proteomic Profiling of Cell Death:Stable Isotope Labeling and Mass Spectrometry Analysis.
Summary
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.
