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Nlrp3

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NLRP3 is a protein that is part of the innate immune system. It functions as a sensor that can detect various stimuli, including pathogens and cellular damage, and triggers an inflammatory response. NLRP3 is a key component of the NLRP3 inflammasome, a protein complex that activates the immune system.

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Lab products found in correlation

Cxcl2, by R&D Systems (2 mentions) Tumor necrosis factor (tnf), by Thermo Fisher Scientific (2 mentions) Il 1β, by Thermo Fisher Scientific (2 mentions) Cxcl1, by R&D Systems (2 mentions) Caspase 1, by Adipogen (2 mentions) Il 18, by R&D Systems (2 mentions) Dnmt3b, by Abcam (1 mentions) Caspase 1, by GeneTex (1 mentions) Alkaline phosphatase conjugated secondary antibody, by Merck Group (1 mentions) Criterion tgx precast protein gel, by Bio-Rad (1 mentions) Annexin a1 anxa1, by Cell Signaling Technology (1 mentions) Il 1β, by LSBio (1 mentions) Immun blot pvdf membrane, by Bio-Rad (1 mentions) Ripa buffer, by Merck Group (1 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions) Halt protease inhibitors cocktail, by Thermo Fisher Scientific (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Prism 8, by GraphPad (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions)

4 protocols using nlrp3

1

Protein Expression Analysis by Western Blot

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Western blots performed with 10, 12, or 15% SDS-polyacrylamide gels were incubated with primary antibodies for IL-1ß (R&D Systems, Minneapolis, MN), NLRP3 (LSBio, Seattle, WA), Ogg1 (Novus Biologicals, Littleton, CO), Caspase 1 (Genetex, Irvine, CA), and DNMT3b (Abcam, Cambridge, MA). Western blots were visualized using alkaline phosphatase-conjugated secondary antibodies (Sigma-Aldrich).
Haldar S., Dru C., Mishra R., Tripathi M., Duong F., Angara B., Fernandez A., Arditi M, & Bhowmick N.A. (2016). Histone deacetylase inhibitors mediate DNA damage repair in ameliorating hemorrhagic cystitis. Scientific Reports, 6, 39257.
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2

Hippocampal Protein and ATP Measurement

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Mouse hippocampi were lysed in radioimmunoprecipitation assay (RIPA) buffer (R0278; Sigma) containing Halt Protease Inhibitors Cocktail (78430; Thermo Scientific). Proteins were separated on 4%-15% or 4%-20% Criterion TGX precast protein gels (64134751; Bio-Rad), and transferred to an Immun-Blot PVDF membrane (1620177; Bio-Rad). After blocking with 5% nonfat dry milk (170-6404; Bio-Rad), the blot was probed with the following antibodies (1:1000): NLRP3 (LS-C374964; LSBio), ASC (67824S; Cell Signaling), Caspase1 (Cleaved Asp210) (LS-C380449; LSBio), IL-1β (LS-C104813; LSBio), and Annexin-A1 (ANXA1) (3299S; Cell Signaling) or (71-3400; Invitrogen). The proteins were visualized using SuperSignal West Dura Extended Duration Substrate (34076; Thermo Scientific) on a ChemiDoc MP Imaging System (Bio-Rad). Protein loading was assessed using anti-GAPDH (2118S; Cell Signaling). Bands were quantified with Fiji software, and analyzed with Prism 8 (GraphPad Software). Hippocampal levels of ATP were determined using a commercially available assay kit (ab83355, Colorimetric/Fluorometric) following the manufacturer’s instructions.
Zhang Z., Ma Q., Velagapudi R., Barclay W.E., Rodriguiz R.M., Wetsel W.C., Yang T., Shinohara M.L, & Terrando N. (2022). Annexin-A1 Tripeptide Attenuates Surgery-Induced Neuroinflammation and Memory Deficits Through Regulation the NLRP3 Inflammasome. Frontiers in Immunology, 13, 856254.
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3

Cytokine and Chemokine Profiling

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Cells were seeded into 6 well plates (106 cells per well), treated as indicated and cell culture supernatants or lavage fluids were analyzed for IL-1β (Invitrogen), IL-18 (R&D), TNF (Invitrogen, R&D Systems), CXCL1 (R&D Systems), CXCL2 (R&D Systems), NLRP3 (LSBio), Caspase-1 (Adipogen) secretion by ELISA as per the manufacturer’s instructions.
Devi S., Indramohan M., Jäger E., Carriere J., Chu L.H., de Almeida L., Greaves D.R., Stehlik C, & Dorfleutner A. (2023). CARD-only proteins regulate in vivo inflammasome responses and ameliorate gout. Cell reports, 42(3), 112265.
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4

Cytokine and Chemokine Profiling

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Cells were seeded into 6 well plates (106 cells per well), treated as indicated and cell culture supernatants or lavage fluids were analyzed for IL-1β (Invitrogen), IL-18 (R&D), TNF (Invitrogen, R&D Systems), CXCL1 (R&D Systems), CXCL2 (R&D Systems), NLRP3 (LSBio), Caspase-1 (Adipogen) secretion by ELISA as per the manufacturer’s instructions.
Japour A.J., Chatis P.A., Eigenrauch H.A, & Crumpacker C.S. (1991). Detection of human immunodeficiency virus type 1 clinical isolates with reduced sensitivity to zidovudine and dideoxyinosine by RNA.RNA hybridization.. Proceedings of the National Academy of Sciences of the United States of America, 88(8), 3092-3096.
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