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Full Description
This second edition provides a sound basis for the molecular investigation of NLR function in health and disease. Chapters focus on NLR as innate immune receptors, "atypical" inflammasomes, biochemical and novel bioluminescence techniques for the measurement of IL-1b as readout of NLR-based inflammasomes, bioluminescent probe, biochemical and microscopy techniques, techniques to measure caspase-1 activation, cell free systems for the study of inflammasome function, and inflammasome activation. 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 key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, NLR Proteins: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researcher sand experts looking to expand their knowledge.
Contents
Canonical Inflammasomes.- Inflammasome-independent roles of NLR and ALR family members.- Measuring IL-1β processing by bioluminescence sensors: using a Bioluminescence Resonance Energy Transfer biosensor.- Methods to measure NLR oligomerization I: size exclusion chromatography, co-immunoprecipitation and cross-linking.- Measuring NLR oligomerization II: detection of ASC speck formation by confocal microscopy and immunofluorescence.- Measuring NLR oligomerization III: detection of NLRP3 and NLRC4 complex by bioluminescence resonance energy transfer.- Method to measure ubiquitinization of NLRs.- Methods to study NLR in human blood cells.- Non-canonical NLRP3 inflammasome activation: standard protocols.- Pyroptosis induction and visualization at the single cell level using optogenetics.- Determination of gasdermin pores.- Methods to activate the NLRP3 inflammasome.- Quantifying cell death induced by the NLRC4 inflammasome.- Methods to activate the NLRP1 inflammasome.- Methods to study inflammasome activation in the central nervous system: immunoblotting and immunohistochemistry.- Inflammasome activation in human macrophages: IL-1β cleavage detection by fully automated capillary-based immunoassay.- Assessing the ATP binding ability of NLRP3 from cell lysates by a pull-down assay.- Theoretical 3D-Modeling of NLRP3 inflammasome complex.- A knock-in mouse model of cryopyrin-associated periodic syndromes.
