Human NLRP3 (NLRP3) Sandwich ELISA (STJE0019810)

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STJE0019810

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Applications: ELISA
Reactivity: Human
Note: FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS.
Sensitivity: 0.09ng/mL
Detection Limit: 0.16~10ng/mL
Short Description: This NLRP3 Sandwich ELISA is an in-vitro enzyme-linked immunosorbent assay for the measurement of samples in human cell culture supernatant, serum and plasma (EDTA, citrate, heparin).
Storage Instruction: If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual.
Assay Time: 3.5h
Detection: Colormetric
Gene Symbol: NLRP3
Gene ID: 114548
Uniprot ID: NLRP3_HUMAN
Specificity: This kit recognizes Human NLRP3 in samples. No significant crossreactivity or interference between Human NLRP3 and analogues was observed.
Sample Type: serum, plasma and other biological fluids
Tissue Specificity Predominantly expressed in macrophages. Also expressed in dendritic cells, B- and T-cells (at protein level). Expressed in LPS-treated granulocytes, but not in resting cells (at protein level). Expression in monocytes is very weak (at protein level). 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). Expressed in lung epithelial cells (at protein level). Expressed in chondrocytes. Expressed at low levels in resting osteoblasts.
Post Translational Modifications Phosphorylation at Ser-198 by MAPK8/JNK1 increases inflammasome activation by promoting deubiquitination by BRCC3 and NLRP3 homooligomerization. Phosphorylation at Ser-806 by CSNK1A1 prevents inflammasome activation by preventing NEK7 recruitment. Phosphorylation at Ser-5 in the pyrin domain inhibits homomultimerization of NLRP3 and activation of the NLRP3 inflammasome: dephosphorylation by protein phosphatase 2A (PP2A) promotes assembly of the NLRP3 inflammasome. Phosphorylation at Ser-295 by PKD/PRKD1 promotes NLRP3 inflammasome assembly. Phosphorylation by ERK1/MAPK3 promotes NLRP3 inflammasome assembly. Phosphorylation by BTK (at Tyr-136, Tyr-140, Tyr-143 and Tyr-168) in the region that mediates binding to phosphatidylinositol phosphate, promotes relocalization of NLRP3 and assembly of the NLRP3 inflammasome. Phosphorylation at Tyr-861 inhibits NLRP3 inflammasome assembly: dephosphorylation by PTPN22 promotes inflammasome activation. Ubiquitinated.undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. Ubiquitination does not lead to degradation, but inhibits inflammasome activation. Deubiquitination is catalyzed by BRCC3 and associated with NLRP3 activation and inflammasome assembly. This process can be induced by the activation of Toll-like receptors (by LPS), through a non-transcriptional pathway dependent on the mitochondrial production of reactive oxygen species, and by ATP. Ubiquitinated by TRIM31 via 'Lys-48'-linked ubiquitination, leading to its degradation by the proteasome. Ubiquitinated at Lys-689 by the SCF(FBXL2) complex, leading to its degradation by the proteasome. Ubiquitinated by TRIM35 via 'lys-48' and 'Lys-63'-linked ubiquitination leading to inhibition of NLRP3 inflammasome activation. Palmitoylation by ZDHHC12 inhibits the NLRP3 inflammasome by promoting NLRP3 degradation by the chaperone-mediated autophagy pathway. Following palmitoylation, HSPA8/HSC70 recognizes and binds the KFERQ-like motifs on NLRP3 and promotes NLRP3 recruitment to lysosomes, where it is degraded via the chaperone-mediated autophagy pathway in a LAMP2-dependent process. Degraded via selective autophagy following interaction with IRGM. IRGM promotes NLRP3 recruitment to autophagosome membranes, promoting its SQSTM1/p62-dependent autophagy-dependent degradation. The disulfide bond in the pyrin domain might play a role in reactive oxygen species-mediated activation. (Microbial infection) ADP-ribosylated by M.pneumoniae CARDS toxin in vitro.
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. 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. 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. Activation of NLRP3 inflammasome is also required for HMGB1 secretion.stimulating inflammatory responses. Under resting conditions, ADP-bound NLRP3 is autoinhibited. 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. Almost all stimuli trigger intracellular K(+) efflux. These stimuli lead to membrane perturbation and activation of NLRP3. 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. Associates with dTGN vesicle membranes by binding to phosphatidylinositol 4-phosphate (PtdIns4P). Shows ATPase activity. Independently of inflammasome activation, regulates the differentiation of T helper 2 (Th2) cells and has a role in Th2 cell-dependent asthma and tumor growth. During Th2 differentiation, required for optimal IRF4 binding to IL4 promoter and for IRF4-dependent IL4 transcription. Binds to the consensus DNA sequence 5'-GRRGGNRGAG-3'. May also participate in the transcription of IL5, IL13, GATA3, CCR3, CCR4 and MAF.
Protein Name Nacht - Lrr And Pyd Domains-Containing Protein 3
Angiotensin/Vasopressin Receptor Aii/Avp-Like
Caterpiller Protein 1.1
Clr1.1
Cold-Induced Autoinflammatory Syndrome 1 Protein
Cryopyrin
Pyrin-Containing Apaf1-Like Protein 1
Database Links Reactome: R-HSA-5689901
Reactome: R-HSA-844456
Reactome: R-HSA-9660826
Reactome: R-HSA-9692916
Reactome: R-HSA-9705671
Reactome: R-HSA-9707564
Cellular Localisation Cytoplasm
Cytosol
Inflammasome
Cytoskeleton
Microtubule Organizing Center
Golgi Apparatus Membrane
Endoplasmic Reticulum
Mitochondrion
Secreted
Nucleus
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
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
Recruited To Dtgn Vesicle Membranes By Binding To Phosphatidylinositol 4-Phosphate (Ptdins4p)
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
Phagocytosis Induces Lysosomal Damage And Inflammasome Activation In The Recipient Cells
In The Th2 Subset Of Cd4(+) Helper T-Cells
Mainly Located In The Nucleus
Nuclear Localization Depends Upon Kpna2
In The Th1 Subset Of Cd4(+) Helper T-Cells
Mainly Cytoplasmic
Alternative ELISA Names Nacht - Lrr And Pyd Domains-Containing Protein 3 ELISA kit
Angiotensin/Vasopressin Receptor Aii/Avp-Like ELISA kit
Caterpiller Protein 1.1 ELISA kit
Clr1.1 ELISA kit
Cold-Induced Autoinflammatory Syndrome 1 Protein ELISA kit
Cryopyrin ELISA kit
Pyrin-Containing Apaf1-Like Protein 1 ELISA kit
NLRP3 ELISA kit
C1orf7 ELISA kit
CIAS1 ELISA kit
NALP3 ELISA kit
PYPAF1 ELISA kit
Specificity This kit recognizes Human NLRP3 in samples. No significant cross-reactivity or interference between Human NLRP3 and analogues was observed.
Reproducibility Both intra-CV and inter-CV are

Information sourced from Uniprot.org


Item Specifications Storage
Micro ELISA Plate (Dismountable) 96T: 8 wells ×12 strips strips -20℃, 6 months
Reference Standard 96T: 2 vials 48T: 1 vial -20℃, 6 months
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL 60 μL -20℃, 6 months
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL 60 μL -20℃ (Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL 2-8°C, 6 months
HRP Conjugate Diluent 1 vial, 14 mL 2-8°C, 6 months
Concentrated Wash Buffer (25×) 1 vial, 30 mL 2-8°C, 6 months
Substrate Reagent 1 vial, 10 mL 2-8℃ (Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 85-96 91
EDTA plasma (n=8) 87-98 92
Cell culture media (n=8) 92-106 98

Intra-assay Precision Intra-assay Precision Intra-assay Precision Inter-assay Precision Inter-assay Precision Inter-assay Precision
Sample 1.00 2.00 3.00 1.00 2.00 3.00
n 20.00 20.00 20.00 20.00 20.00 20.00
Mean (ng/mL) 0.47 0.84 4.71 0.46 0.78 4.71
Standard deviation 0.03 0.04 0.17 0.03 0.04 0.16
CV (%) 6.38 4.76 3.61 6.52 5.13 3.40
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance