Full Description
This volume presents standard approaches and the most recent technical advances used to study innate immune activation. Chapters detail the assessment of macrophage activation, measuring innate immune responses to bacterial viability, quantification of secreted proteins, reporter systems, protocols examining specific innate immune activation by TLRs, RLRs, cGAS, and inflammasomes. 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 practical, Innate Immune Activation: Methods and Protocols aims to be a useful and informative reference tool for further study into this vital field.
Contents
Emerging concepts in innate immunity.- Bioinformatic Assessment of macrophage activation by the innate immune system.- Generation of Genetic Knockouts in myeloid cell lines using a lentiviral CRISPR/Cas9 system.- Modeling primary human monocytes with the trans-differentiation cell line BLaER1.- Measurement of NF-κB activation in TLR-activated macrophages.- Biochemical isolation of the myddosome from murine macrophages.- Generation of innate immune reporter cell lines using retroviral transduction.- Myddosome Formation by Luminescence-based Mammalian Interactome Mapping (LUMIER).- Inflammatory caspases: Activation and cleavage of Gasdermin-D in vitro and during Pyroptosis.- Methods for detecting ASC and ASC speck formation.- Measuring innate immune responses to bacterial viability.- Methods to study cell swelling-induced inflammasome activation.- Detecting release of bacterial dsDNA into the host cytosol using fluorescence microscopy.- Quantitative proteomics of secreted proteins.- Simultaneous detection of cellular viability and interleukin-1β secretion from single cells by ELISpot.- Detection and quantification of MAV aggregation via confocal imaging.
