NLR Proteins : Methods and Protocols / edited by Pablo Pelegrín, Francesco Di Virgilio.

Format:

Book

Contributor:

Pelegrín, Pablo., Editor.

Di Virgilio, Francesco., Editor.

Series:

Methods in Molecular Biology, 1940-6029 ; 2696

Language:

English

Subjects (All):

Proteins.

Local Subjects:

Proteins.

Physical Description:

1 online resource (XII, 299 p. 71 illus., 58 illus. in color.)

Edition:

2nd ed. 2023.

Place of Publication:

New York, NY : Springer US : Imprint: Humana, 2023.

Summary:

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 theirknowledge. .

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.

ISBN:

9781071633502

1071633503