NLRP3 Antibody (N-term)

All lanes : Anti-NLRP3 Antibody (N-term) at 1:1000-1:2000 dilution Lane 1: Hela whole cell lysate Lane 2: Jurkat whole cell lysate Lane 3: Raji whole cell lysate Lane 4: MDA-MB-231 whole cell lysate Lane 5: THP-1 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 118 kDa Blocking/Dilution buffer: 5% NFDM/TBST.
NLRP3 Antibody (N-term) (Cat. #AP8564a) immunohistochemistry analysis in formalin fixed and paraffin embedded human lymph tissue followed by peroxidase conjugation of the secondary antibody and DAB staining. This data demonstrates the use of the NLRP3 Antibody (N-term) for immunohistochemistry. Clinical relevance has not been evaluated.

SPECIFICATION
CITATIONS: 3
PROTOCOLS
BACKGROUND

Product Information
Application Applications Legend: WB=Western Blot IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin-embedded Sections) IHC-F=Immunohistochemistry (Frozen Sections) IF=Immunofluorescence FC=Flow Cytopmetry IC=Immunochemistry ICC=Immunocytochemistry E=ELISA IP=Immunoprecipitation DB=Dot Blot CHIP=Chromatin Immunoprecipitation FA=Fluorescence Assay IEM=Immunoelectronmicroscopy EIA=Enzyme Immunoassay	WB, IHC-P, E
Primary Accession	Q96P20
Reactivity	Human
Host	Rabbit
Clonality	Polyclonal
Isotype	Rabbit IgG
Calculated MW	118173 Da
Antigen Region	65-91 aa

Additional Information
Gene ID	114548
Other Names	NACHT, LRR and PYD domains-containing protein 3, Angiotensin/vasopressin receptor AII/AVP-like, Caterpiller protein 11, CLR11, Cold autoinflammatory syndrome 1 protein, Cryopyrin, PYRIN-containing APAF1-like protein 1, NLRP3, C1orf7, CIAS1, NALP3, PYPAF1
Target/Specificity	This NLRP3 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 65-91 amino acids from the N-terminal region of human NLRP3.
Dilution	WB~~1:1000-1:2000 IHC-P~~1:10~50
Format	Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is purified through a protein A column, followed by peptide affinity purification.
Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Precautions	NLRP3 Antibody (N-term) is for research use only and not for use in diagnostic or therapeutic procedures.

Protein Information
Name	NLRP3 {ECO:0000303\|PubMed:17907925, ECO:0000312\|HGNC:HGNC:16400}
Function	Sensor component of the NLRP3 inflammasome, which mediates inflammasome activation in response to defects in membrane integrity, leading to secretion of inflammatory cytokines IL1B and IL18 and pyroptosis (PubMed:16407889, PubMed:18604214, PubMed:18403674, PubMed:23582325, PubMed:28847925, PubMed:33231615, PubMed:34133077, PubMed:34341353, PubMed:27929086, PubMed:28656979, PubMed:25686105, PubMed:30487600, PubMed:30612879, PubMed:31086327, PubMed:31086329, PubMed:31189953, PubMed:34512673, PubMed:36442502). In response to pathogens and other damage-associated signals that affect the integrity of membranes, initiates the formation of the inflammasome polymeric complex composed of NLRP3, CASP1 and PYCARD/ASC (PubMed:16407889, PubMed:18403674, PubMed:28847925, PubMed:33231615, PubMed:34133077, PubMed:34341353, PubMed:27432880, PubMed:31189953, PubMed:36142182, PubMed:36442502). Recruitment of pro-caspase-1 (proCASP1) to the NLRP3 inflammasome promotes caspase-1 (CASP1) activation, which subsequently cleaves and activates inflammatory cytokines IL1B and IL18 and gasdermin-D (GSDMD), promoting cytokine secretion and pyroptosis (PubMed:23582325, PubMed:28847925, PubMed:33231615, PubMed:34133077, PubMed:34341353, PubMed:31189953). Activation of NLRP3 inflammasome is also required for HMGB1 secretion; stimulating inflammatory responses (PubMed:22801494). Under resting conditions, ADP-bound NLRP3 is autoinhibited (PubMed:35114687). NLRP3 activation stimuli include extracellular ATP, nigericin, reactive oxygen species, crystals of monosodium urate or cholesterol, amyloid-beta fibers, environmental or industrial particles and nanoparticles, such as asbestos, silica, aluminum salts, cytosolic dsRNA, etc (PubMed:16407889, PubMed:18604214, PubMed:18403674, PubMed:19414800, PubMed:23871209). Almost all stimuli trigger intracellular K(+) efflux (By similarity). These stimuli lead to membrane perturbation and activation of NLRP3 (By similarity). Upon activation, NLRP3 is transported to microtubule organizing center (MTOC), where it is unlocked by NEK7, leading to its relocalization to dispersed trans-Golgi network (dTGN) vesicle membranes and formation of an active inflammasome complex (PubMed:36442502). Associates with dTGN vesicle membranes by binding to phosphatidylinositol 4-phosphate (PtdIns4P) (PubMed:30487600, PubMed:34554188). Shows ATPase activity (PubMed:17483456).
Cellular Location	Cytoplasm, cytosol. Inflammasome. Cytoplasm, cytoskeleton, microtubule organizing center. Golgi apparatus membrane. Endoplasmic reticulum {ECO:0000250\|UniProtKB:Q8R4B8}. Mitochondrion. Secreted. Nucleus {ECO:0000250\|UniProtKB:Q8R4B8} Note=In macrophages, under resting conditions, mainly located in the cytosol and on membranes of various organelles, such as endoplasmic reticulum, mitochondria and Golgi: forms an inactive double-ring cage that is primarily localized on membranes (By similarity). Upon activation, NLRP3 is transported to microtubule organizing center (MTOC), where it is unlocked by NEK7, leading to its relocalization to dispersed trans-Golgi network (dTGN) vesicle membranes for the formation of an active inflammasome complex (By similarity). Recruited to dTGN vesicle membranes by binding to phosphatidylinositol 4- phosphate (PtdIns4P) (PubMed:30487600). After the induction of pyroptosis, inflammasome specks are released into the extracellular space where they can further promote IL1B processing and where they can be engulfed by macrophages (PubMed:24952504). Phagocytosis induces lysosomal damage and inflammasome activation in the recipient cells (PubMed:24952504). In the Th2 subset of CD4(+) helper T-cells, mainly located in the nucleus (By similarity). Nuclear localization depends upon KPNA2 (By similarity). In the Th1 subset of CD4(+) helper T-cells, mainly cytoplasmic (By similarity). {ECO:0000250\|UniProtKB:Q8R4B8, ECO:0000269\|PubMed:24952504, ECO:0000269\|PubMed:30487600}
Tissue Location	Predominantly expressed in macrophages (PubMed:33231615, PubMed:34133077). Also expressed in dendritic cells, B- and T-cells (at protein level) (PubMed:11786556) (PubMed:17164409) Expressed in LPS-treated granulocytes, but not in resting cells (at protein level) (PubMed:17164409). Expression in monocytes is very weak (at protein level) (PubMed:17164409). Expressed in stratified non- keratinizing squamous epithelium, including oral, esophageal and ectocervical mucosa and in the Hassall's corpuscles in the thymus Also, detected in the stratified epithelium covering the bladder and ureter (transitional mucosa) (at protein level) (PubMed:17164409) Expressed in lung epithelial cells (at protein level) (PubMed:23229815). Expressed in chondrocytes (PubMed:12032915) Expressed at low levels in resting osteoblasts (PubMed:17907925)

Research Areas

Citations ( 0 )

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

Background

NLRP3 is a pyrin-like protein containing a pyrin domain, a nucleotide-binding site (NBS) domain, and a leucine-rich repeat (LRR) motif. This protein interacts with the apoptosis-associated speck-like protein PYCARD/ASC, which contains a caspase recruitment domain, and is a member of the NALP3 inflammasome complex. This complex functions as an upstream activator of NF-kappaB signaling, and it plays a role in the regulation of inflammation, the immune response, and apoptosis.

References

Feldmann,J., et.al., Am. J. Hum. Genet. 71 (1), 198-203 (2002)

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Affinity Purified Rabbit Polyclonal Antibody (Pab)

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