NLRP3 Rabbit Polyclonal Antibody for WB, IF/ICC, ELISA - Q96P20

Core Product Specifications and Parameters

Biological Background: NLRP3 Function and Localization

• NLRP3 (NACHT, LRR and PYD domains-containing protein 3), also known as cryopyrin, CIAS1, or NALP3, is a member of the NOD-like receptor (NLR) family, characterized by an N-terminal pyrin domain (PYD), a central NACHT domain, and C-terminal leucine-rich repeats (LRRs).
• As the sensor component of the NLRP3 inflammasome, it orchestrates the recruitment of ASC (PYCARD) and pro-caspase-1, leading to caspase-1 activation, which cleaves pro-IL-1β and pro-IL-18 into mature cytokines and triggers gasdermin-D-mediated pyroptosis.
  Related references: PMID:16407889 PMID:18403674 PMID:18604214 PMID:27432880 PMID:23582325 PMID:28847925 PMID:31189953 PMID:22801494
• NLRP3 responds to a broad spectrum of stimuli, including extracellular ATP, nigericin, monosodium urate crystals, cholesterol crystals, amyloid-β fibrils, and environmental particulates; these signals induce potassium efflux and membrane perturbation, resulting in NLRP3 translocation to the microtubule organizing center (MTOC) and dispersed trans-Golgi network (dTGN), where it binds phosphatidylinositol 4-phosphate (PtdIns4P) via its polybasic region, facilitating inflammasome assembly.
  Related references: PMID:16407889 PMID:18403674 PMID:18604214 PMID:36442502 PMID:39173637 PMID:30487600 PMID:34554188
• Predominantly expressed in macrophages, and also detected in dendritic cells, B- and T-cells, granulocytes, lung epithelial cells, chondrocytes, and stratified squamous epithelia (oral, esophageal, ectocervical, and bladder).
  Related references: PMID:33231615 PMID:34133077 PMID:11786556 PMID:17164409 PMID:23229815 PMID:12032915
• Gain-of-function mutations in NLRP3 are responsible for cryopyrin-associated periodic syndromes (CAPS), encompassing familial cold autoinflammatory syndrome, Muckle-Wells syndrome, and CINCA/NOMID, and are associated with deafness and systemic amyloidosis.
• Subcellular localization includes the cytosol, inflammasome complex, Golgi apparatus membrane, endoplasmic reticulum, mitochondria, and it can be secreted or found in the nucleus under certain conditions.

Experimental Guidance and Technical Tips

• The immunogen spans amino acids 100–210, which lies within the PYD domain of human NLRP3; this region is relatively conserved across species, consistent with the observed cross-reactivity with mouse and rat orthologs.
• Western blot is expected to show a single band at approximately 118 kDa in total cell lysates from macrophages or other NLRP3-expressing cells.
• For positive controls, consider using LPS‑treated human THP‑1 cells or mouse bone marrow‑derived macrophages, known to upregulate NLRP3.
• In IF/ICC, permeabilization and fixation conditions may need optimization; including unstimulated vs. stimulated samples can help visualize inflammasome speck formation.
• Validate antibody performance in your specific sample type and application, as the polyclonal nature may lead to batch‑to‑batch variability; pilot tests with proper controls are recommended.

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