WB
Western blot analysis of NLRP3 on different lysates with Rabbit anti-NLRP3 antibody at 1/1,000 dilution. Lane 1: THP-1 cell lysate, Lane 2: RAW264.7 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-NLRP3 antibody at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells treated with 10μg/mL LPS for 8 hours labeling NLRP3 with Rabbit anti-NLRP3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NLRP3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of NLRP3 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).WB
(E-F) Western blot results of NLRP3, ER, Wnt2b, and β-catenin different cells.| Product Name | NLRP3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within Human NLRP3 aa 5-161 / 1036. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:500-1:1000 IHC:1:100-1:400 ICC:1:100-1:500 FC:1:50-1:100 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NLRP3 |
|---|---|
| Gene Synonyms | AII AVP FCU MWS FCAS KEFH CIAS1 FCAS1 NALP3 C1orf7 CLR1.1 DFNA34 PYPAF1 AGTAVPRL |
| Gene Full Name | NLR family pyrin domain containing 3 |
| Gene Summary | This gene encodes 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 NLRP3 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. The SARS-CoV 3a protein, a transmembrane pore-forming viroporin, has been shown to activate the NLRP3 inflammasome via the formation of ion channels in macrophages. Mutations in this gene are associated with familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous and articular (CINCA) syndrome, neonatal-onset multisystem inflammatory disease (NOMID), keratoendotheliitis fugax hereditarian, and deafness, autosomal dominant 34, with or without inflammation. Multiple alternatively spliced transcript variants encoding distinct isoforms have been identified for this gene. Alternative 5' UTR structures are suggested by available data; however, insufficient evidence is available to determine if all of the represented 5' UTR splice patterns are biologically valid. [provided by RefSeq, Aug 2020] |
| Alternative Names | AGTAVPRL antibody AII/AVP antibody Angiotensin/vasopressin receptor AII/AVP like antibody Angiotensin/vasopressin receptor AII/AVP-like antibody C1orf7 antibody Caterpiller protein 1.1 antibody CIAS 1 antibody CIAS1 antibody CLR1.1 antibody Cold autoinflammatory syndrome 1 antibody AGTAVPRL antibody AII/AVP antibody Angiotensin/vasopressin receptor AII/AVP like antibody Angiotensin/vasopressin receptor AII/AVP-like antibody C1orf7 antibody Caterpiller protein 1.1 antibody CIAS 1 antibody CIAS1 antibody CLR1.1 antibody Cold autoinflammatory syndrome 1 antibody Cold autoinflammatory syndrome 1 protein antibody Cryopyrin antibody Familial cold autoinflammatory syndrome antibody FCAS antibody FCU antibody LRR and PYD domains-containing protein 3 antibody Muckle-Wells syndrome antibody MWS antibody NACHT antibody NACHT LRR and PYD containing protein 3 antibody NALP 3 antibody NALP3 antibody NALP3_HUMAN antibody NLRP3 antibody PYPAF 1 antibody PYPAF1 antibody PYRIN containing APAF1 like protein 1 antibody PYRIN-containing APAF1-like protein 1 antibody |
| Molecular Weight(MW) | 118kDa |
| Cellular Localization | Cytoplasm, Inflammasome, Secreted, Nucleus, Endoplasmic reticulum. |
WB
Western blot analysis of NLRP3 on different lysates with Rabbit anti-NLRP3 antibody at 1/1,000 dilution. Lane 1: THP-1 cell lysate, Lane 2: RAW264.7 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-NLRP3 antibody at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells treated with 10μg/mL LPS for 8 hours labeling NLRP3 with Rabbit anti-NLRP3 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NLRP3 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of NLRP3 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
WB
(E-F) Western blot results of NLRP3, ER, Wnt2b, and β-catenin different cells.| Application Notes | WB:1:500-1:1000 IHC:1:100-1:400 ICC:1:100-1:500 FC:1:50-1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
