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Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
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Immunology and Infection
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JoVE Journal Immunology and Infection
Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells

English

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20,633 Views

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09:04 min

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May 22, 2014

DOI:

10.3791/51284-v

DOI:
10.3791/51284-v

•

09:04 min
May 22, 2014

•

4 Views
, ,
1Department of Pathology,New York University School of Medicine, 2Division of Infectious Diseases, Department of Medicine,Mount Sinai Medical Center, 3Division of Hematology and Oncology, Hess Center for Science and Medicine,Mount Sinai Medical Center

Summary

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Dendritic cells (DCs) secrete IL-1β in response to TLR8 recognition of synthetic purine, R848, followed by NLRP3 inflammasome activation with nigericin, therefore, IL-1β can be used to measure NLRP3 inflammasome activity. Intracellular cytokine staining, immunoblotting, and ELISA are used to accurately measure NLRP3 inflammasome priming and activation via IL-1β expression.

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Cite this Article

Fernandez, M. V., Miller, E. A., Bhardwaj, N. Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells. J. Vis. Exp. (87), e51284, doi:10.3791/51284 (2014).

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