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This volume explores the latest techniques and approaches used to study this emerging form of cell death. The chapters in this book cover topics such as distinguishing necroptosis from apoptosis; detecting MLKL oligomerization during programmed necrosis; analyzing the RIPK3 necrosome complex; using RIPK1 kinase small molecule inhibitors in studying necroptosis; monitoring RIPK1 phosphorylation in the TNFR1 signaling complex; and assessing in vivo kidney cell death in glomerular injury. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Programmed Necrosis: Methods and Protocols is a valuable resource for technicians, graduate students, postdoctoral fellows, and other scientists who are pursuing studies in this field.
List of contents
Tools in the Art of Studying Necroptosis.- Loss-of-Function RNAi Screen to Identify Necrosis-Signaling Molecules.- Chemical Library Screens to Identify Pharmacological Modulators of Necroptosis.- Distinguishing Necroptosis from Apoptosis.- Methods for Studying TNF-Mediated Necroptosis in Cultured Cells.- Analysis of Necroptosis in Bone Marrow-Derived Macrophages.- Generation and Use of Chimeric RIP Kinase Molecules to Study Necroptosis.- Detection of MLKL Oligomerization during Programmed Necrosis.- Analysis of Cytokine- and Influenza A Virus-Driven RIPK3 Necrosome Formation.- Detection of RIPK1 in the FADD-Containing Death Inducing Signaling Complex (DISC) during Necroptosis.- Use of RIP1 Kinase Small Molecule Inhibitors in Studying Necroptosis.- Analyzing Necroptosis using a RIPK1 Kinase Inactive Mouse Model of TNF Shock.- Assessment of In Vivo Kidney Cell Death: Acute Kidney Injury.- Assessment of In Vivo Kidney Cell Death: Glomerular Injury.- Detection of Necroptosis by Phospho-RIPK3 Immunohistochemical Labeling.- Characterization of the TNFR1-SC using 'Modified Tandem Affinity Purification' in Conjunction with Liquid Chromatography-Mass Spectrometry (LC-MS).- Monitoring RIPK1 Phosphorylation in the TNFR1 Signaling Complex.- Analysis of CYLD Proteolysis by Caspase-8 in Bone Marrow-Derived Macrophages.
