The Inflammasome, 1st ed. 2022 Methods and Protocols Methods in Molecular Biology Series, Vol. 2459

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.

1. Assessment of ASC Oligomerization by Flow Cytometry

Laura Hurtado-Navarro, Alberto Baroja-Mazo, Helios Martínez-Banaclocha, and Pablo Pelegrín

2.In-vitro Assays to Study Inflammasome Activation in Primary Macrophages

Ishita Banerjee

3. Measuring Non-Canonical Inflammasome Activation In Neutrophils

Mercedes Monteleone and Dave Boucher

Louisa Janice Kamajayaand Dave Boucher

5. Activation of the Non-Canonical Inflammasome In Mouse And Human Cells

Jelena S. Bezbradica, Rebecca C. Coll, and Dave Boucher

6. Measurement of Inflammasome-Induced Mitochondrial Dysfunction By Flow Cytometry

Mayoorey Murugathasan and Ali A. Abdul-Sater

7. Detection of ASC Oligomerization by Western Blotting

Safoura Zangiabadi, Ali Akram, and Ali A. Abdul-Sater

8. Reconstitution System of NLRP3 Inflammasome in HEK293T Cells

Sheng Chen, Zhexu Chi, and Di Wang

9. Intracellular Potassium Ions Measurements by Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES)

Yifei Zhangand Yan Shi

10. NLRP3 Phospho-Residue Mapping by Phospho Dot-Blots

Sangeetha Shankar, Zsofia Bittner, Alexander N. R. Weber

11. Analysis of Activity and Expression of the NLRP3, AIM2 and NLRC4 Inflammasome in Whole Blood

Lev Grinstein and Stefan Winkler

12. Measurement of Cytosolic Mitochondrial DNA after NLRP3 Inflammasome Activation

Olga M. Antónand Javier Traba

13. Inflammasome-derived Exosomes Isolation and Imaging by Transmission Electron Microscopy

Yuehui Zhang and Jian Wang

14. In Vitro and In Vivo Model for Sepsis through Non-Canonical NLRP3 inflammasome Activation

Xiao Sun, Tae-Bong Kang, and Kwang-Ho Lee

15. Inflammasome Activation in Gingival Epithelial Cells

Kalina R Atanasovaand Özlem Yilmaz

Lucas Secchim Ribeiro

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts