Measuring the Inflammasome
Inflammasomes are multiprotein complexes whose activity has been implicated in physiological and pathological inflammation. The hallmarks of inflammasome activation are the secretion of the mature forms of Caspase-1 and IL-1β from cells of the innate immune system. This protocol covers the methods required to study inflammasome activation using mouse bone marrow-derived dendritic cells (BMDCs) as a model system. The protocol includes the generation and handling of BMDCs, the stimulation of BMDCs with established Nlrp3 inflammasome activators, and the measurement of activation by both ELISA and western blot. These methods can be useful for the study of potential inflammasome activators, and of the signaling pathways involved in inflammasome activation. General considerations are provided that may help in the design and optimization of modified methods for the study of other types of inflammasomes and in other cell types.
Key wordsInflammasome Nlrp3 Caspase-1 Interleukin-1 Bone marrow-derived dendritic cells
I would like to thank Michael Bscheider, Manuel Ritter, and James Vince for helpful suggestions, and Christina Thomas for proofreading and correcting the manuscript. My work is funded by an EMBO long-term fellowship. I declare no competing financial interests.
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