- MA5-23919 - Invitrogen Antibodies NLRP3 antibody

Submitted references Modified Citrus Pectin Alleviates Cerebral Ischemia/Reperfusion Injury by Inhibiting NLRP3 Inflammasome Activation via TLR4/NF-ĸB Signaling Pathway in Microglia.

Cui Y, Zhang NN, Wang D, Meng WH, Chen HS
Journal of inflammation research 2022;15:3369-3385

miR-25-3p ameliorates SAE by targeting the TLR4/NLRP3 axis.

Luo XY, Ying JH, Wang QS
Metabolic brain disease 2022 Aug;37(6):1803-1813

Huaier extract suppresses non-small cell lung cancer progression through activating NLRP3-dependent pyroptosis.

Xie J, Zhuan B, Wang H, Wang Y, Wang X, Yuan Q, Yang Z
Anatomical record (Hoboken, N.J. : 2007) 2021 Feb;304(2):291-301

Sauchinone Protects Renal Mesangial Cell Dysfunction against Angiotensin II by Improving Renal Fibrosis and Inflammation.

Yoon JJ, Lee HK, Kim HY, Han BH, Lee HS, Lee YJ, Kang DG
International journal of molecular sciences 2020 Sep 23;21(19)

Hyperactivity of Innate Immunity Triggers Pain via TLR2-IL-33-Mediated Neuroimmune Crosstalk.

Huang J, Gandini MA, Chen L, M'Dahoma S, Stemkowski PL, Chung H, Muruve DA, Zamponi GW
Cell reports 2020 Oct 6;33(1):108233

Microbial BMAA elicits mitochondrial dysfunction, innate immunity activation, and Alzheimer's disease features in cortical neurons.

Silva DF, Candeias E, Esteves AR, Magalhães JD, Ferreira IL, Nunes-Costa D, Rego AC, Empadinhas N, Cardoso SM
Journal of neuroinflammation 2020 Nov 5;17(1):332

Extracellular ASC exacerbated the recurrent ischemic stroke in an NLRP3-dependent manner.

He XF, Zeng YX, Li G, Feng YK, Wu C, Liang FY, Zhang Y, Lan Y, Xu GQ, Pei Z
Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 2020 May;40(5):1048-1060

Monitoring Inflammasome Priming and Activation in Response to Candida albicans.

Santana DJ, Anderson FM, O'Meara TR
Current protocols in microbiology 2020 Dec;59(1):e124

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis from lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line and RAW 264.7 mouse monocyte/macrophage cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-human/mouse NLRP3/NALP3 Monoclonal Antibody (Product # MA5-23919) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody. A specific band was detected for NLRP3/NALP3 at approximately 117 kDa (as indicated). This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of NLRP3 in J774A.1 mouse reticulum cell sarcoma macrophage cell line and RAW 264.7 mouse monocyte/macrophage cell line. Samples were incubated in NLRP3 monoclonal antibody (Product # MA5-23919) using a dilution of 2 µg/mL followed by a HRP-conjugated Anti-Rat IgG secondary antibody. A specific band was detected for NLRP3/NALP3 at approximately 117 kDa (as indicated). This experiment was conducted under reducing condition. Recommended for use with human lysates.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry analysis of NLRP3 in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line. Samples were incubated in NLRP3 monoclonal antibody (Product # MA5-23919) using a dilution of 10 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of NLRP3 in immersion fixed paraffin-embedded sections of human tonsil tissue. Samples were incubated in NLRP3 monoclonal antibody (Product # MA5-23919) using a dilution of 5 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic . Tissue was stained using the Anti-Rat IgG VisUCyte™ HRP Polymer Antibody (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometric analysis of human peripheral blood monocytes was stained with Rat Anti-human/mouse NLRP3/NALP3 Monoclonal Antibody (Product # MA5-23919) or isotype control antibodyopen histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of human peripheral blood monocytes was stained with Rat Anti-human/mouse NLRP3/NALP3 Monoclonal Antibody (Product # MA5-23919) or isotype control antibodyopen histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of NLRP3 in RAW 264.7 mouse monocyte/macrophage cell line. Samples were incubated in NLRP3 monoclonal antibody (Product # MA5-23919) or isotype control antibody followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of NLRP3 in Human peripheral blood monocytes. Samples were incubated in NLRP3 monoclonal antibody (Product # MA5-23919) or isotype control antibody followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Inhibition effects of sauchinone on the NOD-like receptor family, pyrin domain-containing-3 (NLRP3) inflammasome in mesangial cells. ( A ) Western blotting and ( B ) real-time PCR showed protein and mRNA levels of NLRP3, ASC, IL-1beta, cleaved caspase-1, and caspase-1 in sauchinone-treated and AngII-stimulated cells at 48 h. beta-actin or GAPDH were used as the internal standard in each sample. ** p < 0.01 vs. control and ## p < 0.01 vs. AngII alone.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 BMAA activates neuronal innate immunity. In primary mouse cortical neurons pre-treated with 1 muM CCCP, 3 mM BMAA, and 1 mug/mul LPS for 48 h or with 3 mM BMAA for6, 24 and 48 h, we evaluated a changes in protein levels of TLR4 and b TLR 3 by western blotting. Representative immunoblot and densitometric analysis was performed by the loading of equal amounts of protein corrected with beta-actin. Inflammasome activation was determined by the quantification of c cytosolic IkBalpha protein levels which were determined by western blotting. Representative immunoblot and densitometric analysis was performed by loading equal amounts of protein corrected with beta-actin. d p65 NFkappaB was detected in enriched nuclear fractions by western blotting. Representative immunoblot and densitometric analysis was performed by the loading of equal amounts of protein corrected with TATA-binding protein. e Cytosolic NLRP3 protein levels were evaluated by western blotting. Representative immunoblot and densitometric analysis was performed by loading equal amounts of protein corrected against beta-actin. f Cytosolic pro-IL-1beta protein levels analyzed by western blotting. Representative immunoblot and densitometric analysis was performed by the loading of equal amounts of protein corrected with beta-actin. g p62 flux was evaluated by western blotting after 4-h incubation with or without NH 4 Cl plus leupeptin. Representative immunoblot and densitometric analysis was performed by the loading

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 1 FIGURE Pyroptosis levels were decreased in non-small cell lung cancer (NSCLC) tissues and cell lines. (a and c) The mRNA levels of pyroptotic-related genes (NLRP3, caspase-1, IL-1beta, and IL-18) in NSCLC tissues, adjacent noncancerous tissues, NSCLC cell lines (A549, H358, and H520), and normal lung epithelial cell line BEAS-2B were analyzed by using qRT-PCR. (b and d) The proteins levels of pyroptotic-related genes (NLRP3, caspase-1, IL-1beta, and IL-18) in NSCLC tissues, adjacent noncancerous tissues, NSCLC cell lines (A549, H358, and H520), and normal lung epithelial cell line BEAS-2B were analyzed by using western blot. GAPDH is chosen as protein controls. ** p < .01

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 6 FIGURE Huaier suppresses tumor growth and induces pyroptosis in vivo. Tumorigenesis assay was performed by subcutaneous injection of H520 cells into flanks of BALB/c nude mice. (a) Tumor volumes were measured at the indicated days post-injection. (b) The effect of huaier on apoptosis in tumor tissues was evaluated by TUNEL assay. (c) The western blot analysis detected the relative expression of Ki-67, NLRP3, caspase-1, IL-1beta, and IL-18 in tumor tissues. (d) Bodyweight of mice in the vehicle group, low dose group (50 mg/ml), and high dose group (100 mg/ml) were evaluated every 3 days. (e) H&E staining was applied to evaluate the changes in the heart, liver, spleen, lung, and kidney tissues. ** p < .01

MA5-23919

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