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This volume provides inflammasome instructions and tips to study different aspects of inflammasome biology. Chapters guide readers through how to assess canonical and non-canonical inflammasome activation, in vitro immune and non-immune cells, whole blood, biochemical assays to evaluate ASC oligomerization, gasdermin D cleavage, and mapping phosphorylated residues of NLRP3. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, The Inflammasome: Methods and Protocols aims to provide protocols that will serve as tools for current and aspiring researchers interested in studying inflammasomes.
List of contents
Assessment of ASC Oligomerization by Flow Cytometry.- In-vitro Assays to Study Inflammasome Activation in Primary Macrophages.- Measuring Non-Canonical Inflammasome Activation In Neutrophils.- Gasdermin D Cleavage Assay Following Inflammasome Activation.- Activation of the Non-Canonical Inflammasome In Mouse And Human Cells.- Measurement of Inflammasome-Induced Mitochondrial Dysfunction By Flow Cytometry.- Detection of ASC Oligomerization by Western Blotting.- Reconstitution System of NLRP3 Inflammasome in HEK293T Cells.- Intracellular Potassium Ions Measurements by Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES).- NLRP3 Phospho-Residue Mapping by Phospho Dot-Blots.- Analysis of Activity and Expression of the NLRP3, AIM2 and NLRC4 Inflammasome in Whole Blood.- Measurement of Cytosolic Mitochondrial DNA after NLRP3 Inflammasome Activation.- Inflammasome-derived Exosomes Isolation and Imaging by Transmission Electron Microscopy.- In Vitro and In Vivo Model for Sepsis through Non-Canonical NLRP3 inflammasome Activation.- Inflammasome Activation in Gingival Epithelial Cells.- Imaging of Inflammasome Speck Formation in Living Cells.
