NLR Proteins Methods and Protocols

This volume provides a sound basis for the molecular investigation of NLR function in health and disease. Chapters focus on of innate immune receptors, “atypical” inflammasomes, biochemical and novel bioluminescence techniques for the measurement of IL-1b, 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 tips on troubleshooting and avoiding known pitfalls.

 

Authoritative and cutting-edge, NLR Proteins: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Innate Immune Receptors.- Atypical Inflammasomes.- Assessment of Inflammasome Activation By Cytokine And Danger Signal Detection.- Investigating IL-1ß Secretion Using Real-Time Single-Cell Imaging.- Measuring IL-1ß Processing By Bioluminescence Sensors I: Using a Bioluminescence Resonance Energy Transfer Biosensor.- Measuring IL-1ß Processing By Bioluminescence Sensors II: The Igluc System.- Assessing Extracellular ATP as Danger Signal In Vivo: The Pmeluc System.- Measuring 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 Complex by Bioluminescence Resonance Energy Transfer.- Measuring NLR Oligomerization IV: Using Förster Resonance Energy Transfer (FRET)-Fluorescence Lifetime Imaging Microscopy (FLIM) to Determine the Close Proximity Of Inflammasome Components.- Measuring NLR Oligomerization V: In Situ Proximity Ligation Assay.- Assessing Caspase-1 Activation.- Cell-Free Assay for Inflammasome Activation.- Functional Reconstruction of Nlrs In HEK293 Cells.- Method to Measure Ubiquitinization of Nlrs.- Cytofluorometric Quantification of Cell Death Elicited By NLR Proteins.- NLR in Human Diseases: Role and Laboratory Findings.