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Intracellular NLRP3 Assay
Cell-Based Assays

Intracellular NLRP3 Assay Services

NLRP3 is an innate immune receptor protein that acts as a molecular sensor within cells. The NLRP3 inflammasome is a cytosolic multiprotein complex composed of NLRP3, adapter protein ASC, and inflammatory protease caspase-1 that responds to microbial infection, endogenous danger signals, and environmental stimuli. The assembled NLRP3 inflammasome activates the protease caspase-1 to induce pyroptosis and facilitate the release of IL-1β and IL-18, which contribute to innate immune defense and homeostatic maintenance.

NanoBRET Target Engagement Assay for NLRP3 in Live Cells

NanoBRET Target Engagement (TE) is a biophysical method of assessing the binding between a compound and a target protein in live cells. Based on bioluminescent resonance energy transfer (BRET), NanoBRET TE can be applied to the innate immune receptor protein NLRP3, which is involved in both innate immune responses and the development of various diseases, including autoinflammatory conditions, autoimmune diseases, and neurodegenerative disorders.

List of Targets

Assay Details

  • Assay Format
  • Screening Details
Assay Format

In the NanoBRET TE Assay for NLRP3, the target NLRP3 protein is expressed as a NanoLuc Luciferase fusion. A cell-permeable, fluorescent NanoBRET NLRP3 tracer is added. The tracer reversibly binds to the NLRP3-NanoLuc fusion protein. When in proximity to one another, the tracer and NanoLuc Luciferase produce a BRET signal. When a test compound binds to NLRP3, it competes with the tracer, and the BRET signal is lost.

Screening Details

Setups: IC50 value determination in 10-dose duplicate assay format.

Controls: We use DMSO as a negative control. In every project, testing of a standard reference compound (MCC950) is included in IC50 assay format free of charge.

Turnaround time: 2-3 weeks. Expedited scheduling and data delivery can be arranged prior to study commencement.

Report: A detailed report including assay conditions and comprehensive evaluation of data as well as raw data for each analysis will be provided.

Screening location: Malvern, PA, USA

Compound requirements: In brief, 50 to 100 µl of a 10 to 50 mM DMSO stock or 2-3 mg solid material is needed. Please refer to our FAQs for information regarding compound preparation and shipping.