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Immunology and Infection

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

20.7K Views

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09:04 min

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May 22nd, 2014

DOI :

10.3791/51284-v

May 22nd, 2014

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Melissa V. Fernandez¹, Elizabeth A. Miller², Nina Bhardwaj³

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

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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Chapters in this video

0:05

Title

1:34

Priming and Activating the Inflammasome

2:46

Sample Collection

5:05

Data Acquisition

6:52

Results: Measuring TLR8 Priming with R848

8:33

Conclusion

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