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This volume provides detailed protocols for the performance, analysis, and troubleshooting of in vitro and in vivo experiments related to programmed cell death. Chapters compile conventional techniques such as western blot and qPCR and state-of-the art transmission electron microscopy and real-time multiplexed imaging assays. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Detection of Cell Death Mechanisms: Methods and Protocols aims to ensure seamless execution of protocols on specific cell death type.
List of contents
Determination of Caspase Activation by Western Blot.- Detection of Unfolded Protein Response by Polymerase Chain Reaction.- Subcellular Fractionation to Demonstrate Activation of Intrinsic Apoptotic Pathway.- A Triple-Parameter Based Laboratory Friendly Fluorescence Imaging to Identify Apoptosis in Live Cells.- Flow Cytometric Analyses of p53-mediated Cell Cycle Arrest and Apoptosis in Cancer Cells.- A Real-Time, Bioluminescent Apoptosis Assay.- Detection of Anoikis Using Cell Viability Dye and Quantitation of Caspase Activity.- Time and Dose Dependent Toxicity Studies in 3D Cultures Using a Luminescent Lactate Dehydrogenase Assay.- Visualization and Quantification of Neutrophil Extracellular Traps.- 2In Vitro Identification and Isolation of Human Neutrophil Extracellular Traps.- Induction and Detection of Necroptotic Cell Death in Mammalian Cell Culture.- Visualization of Necroptotic Cell Death through Transmission Electron Microscopy.- Methodology for Comprehensive Detection of Pyroptosis.- Determination and Quantitation of Cytotoxic T Cell-Mediated Cell Death.- Detection of Immunogenic Cell Death in Tumor Vaccination Mouse Model.- A Luminescent Assay to Quantify Cell Viability in Real Time.- Kinetic Measurement of Apoptosis and Immune Cell Killing Using Live-Cell Imaging and Analysis.- Two Plasmid-Based Systems for CRISPR/Cas9 Mediated Knockout of Target Genes.- Determining Cell Death Pathway and Regulation by Enrichment Analysis.- Transcription Factor Binding Site Identification and Enrichment Analysis.
