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Tlrl msu

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Tlrl-msu is a synthetic ligand for the toll-like receptor 3 (TLR3). It functions as a potent and selective agonist for TLR3, which is involved in the recognition of viral double-stranded RNA and the subsequent activation of the innate immune response.

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Lab products found in correlation

Human ifn β, by R&D Systems (2 mentions) Mouse ifn β, by R&D Systems (2 mentions) Mouse il 18, by R&D Systems (2 mentions) Human il 18, by R&D Systems (2 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions) Tlrl 3pelps, by InvivoGen (2 mentions) Tlrl atp, by InvivoGen (2 mentions) Hmgb1, by Thermo Fisher Scientific (1 mentions) Pam3csk4, by Thermo Fisher Scientific (1 mentions) Fsl 1, by InvivoGen (1 mentions) Imiquimod, by InvivoGen (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Trizol, by Merck Group (1 mentions) Tnf α, by R&D Systems (1 mentions) Ifn γ, by R&D Systems (1 mentions) Tlrl nig, by InvivoGen (1 mentions) Lipofectamine, by InvivoGen (1 mentions) Tlrl mdpc, by InvivoGen (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions)

Close spelling variants of this product

MSU (tlrl‐msu)

5 protocols using tlrl msu

1

Induction of Caspase-11 in Bone Marrow-Derived Macrophages

Cited 2 times
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Bone marrow-derived macrophages (BMDMs) were cultured as previously described (25 (link), 80 (link), 81 (link), 83 (link)–85 (link)). Caspase-11 induction was measured by treating WT macrophages with various inflammatory mediators for 4 h. 20 ng/ml IL-1α, IL-1β, IL-18, and KC (R&D Systems, 400-ML-005, 401-ML-005, 9139-IL-050, and 453-KC-050), HMGB1 (eBioscience, 34-8401-82), 2.5 μg/ml Pam3CSK4 (tlrl-pms), 5 μg/ml poly(I:C) (tlrl-picw), 100 ng/ml LPS (tlrl-eklps), 500 ng/ml flagellin (tlrl-stfla), 100 ng/ml FSL-1 (tlrl-fsl), 5 μg/ml ssRNA (tlrl-lrna40), 5 μg/ml Imiquimod (tlrl-imqs), and 100 ng/ml bacterial DNA (tlrl-ssec) (InvivoGen). Interferon induction of caspase-11 was achieved using 200 ng/ml IFNα and β (BioLegend, 752806, 581306), 50 ng/ml IFNγ (R&D Systems, 485-MI), and 100 ng/ml TNFα (R&D Systems, 410-MT-050) and LPS for 4 h, respectively. In vitro stimulation with MSU was accomplished with 100 μg/ml (Invivogen, tlrl-msu). ATP (Sigma-Aldrich) was used for 30 min at the final concentration of 5 mM. All cells were lysed and stored in TRIzol™.
Caution K., Young N., Robledo-Avila F., Krause K., Abu Khweek A., Hamilton K., Badr A., Vaidya A., Daily K., Gosu H., Anne M.N., Eltobgy M., Dakhlallah D., Argwal S., Estfanous S., Zhang X., Partida-Sanchez S., Gavrilin M.A., Jarjour W.N, & Amer A.O. (2019). Caspase-11 Mediates Neutrophil Chemotaxis and Extracellular Trap Formation During Acute Gouty Arthritis Through Alteration of Cofilin Phosphorylation. Frontiers in Immunology, 10, 2519.
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2

Cytokine Secretion Assay in Mouse RPE Cells

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Secreted human and mouse interferon-β and IL-18 in the media were detected using ELISA kits (mouse IFN-β, R&D Systems Cat# 42400-1; mouse IL-18, R&D Systems Cat# 7625; human IFN-β, R&D Systems Cat# 41410; human IL-18, R&D systems Cat# DY318-05) according to the manufacturer′s instructions. Primary mouse cells were cultured as above. WT, Gsdmd−/−, Casp11−/−, or Mb21d1−/− mouse RPE cells were seeded at a density of 250,000 cells/well in a 12-well plate. When confluency reached 60–70%, cells were transfected with 20 pmol of in vitro transcribed Alu RNA or mock using Lipofectamine 2000 reagent (Life Technologies, Carlsbad, CA) following the manufacturer’s protocol. Media was collected to detect secreted cytokine content at 8 to 24 h post-transfection. For examination of the induction of IL-18 secretion by monosodium urate (MSU) crystals (Invivogen Cat# tlrl-msu), mouse RPE cells were primed with LPS (500 ng/ml) for 6 h and exposed to MSU (250 μg/ml) for 16 h, and media was collected to detect secreted cytokine.
Kerur N., Fukuda S., Banerjee D., Kim Y., Fu D., Apicella I., Varshney A., Yasuma R., Fowler B.J., Baghdasaryan E., Marion K.M., Huang X., Yasuma T., Hirano Y., Serbulea V., Ambati M., Ambati V.L., Kajiwara Y., Ambati K., Bastos-Carvalho A., Ogura Y., Terasaki H., Oshika T., Kim K.B., Hinton D.R., Leitinger N., Cambier J.C., Buxbaum J.D., Kenney M.C., Jazwinski S.M., Nagai H., Hara I., West A.P., Fitzgerald K.A., Sadda S.R., Gelfand B.D, & Ambati J. (2017). cGAS drives non-canonical inflammasome activation in age-related macular degeneration. Nature medicine, 24(1), 50-61.
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3

Cytokine Secretion Assay in Mouse RPE Cells

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Secreted human and mouse interferon-β and IL-18 in the media were detected using ELISA kits (mouse IFN-β, R&D Systems Cat# 42400-1; mouse IL-18, R&D Systems Cat# 7625; human IFN-β, R&D Systems Cat# 41410; human IL-18, R&D systems Cat# DY318-05) according to the manufacturer′s instructions. Primary mouse cells were cultured as above. WT, Gsdmd−/−, Casp11−/−, or Mb21d1−/− mouse RPE cells were seeded at a density of 250,000 cells/well in a 12-well plate. When confluency reached 60–70%, cells were transfected with 20 pmol of in vitro transcribed Alu RNA or mock using Lipofectamine 2000 reagent (Life Technologies, Carlsbad, CA) following the manufacturer’s protocol. Media was collected to detect secreted cytokine content at 8 to 24 h post-transfection. For examination of the induction of IL-18 secretion by monosodium urate (MSU) crystals (Invivogen Cat# tlrl-msu), mouse RPE cells were primed with LPS (500 ng/ml) for 6 h and exposed to MSU (250 μg/ml) for 16 h, and media was collected to detect secreted cytokine.
Kerur N., Fukuda S., Banerjee D., Kim Y., Fu D., Apicella I., Varshney A., Yasuma R., Fowler B.J., Baghdasaryan E., Marion K.M., Huang X., Yasuma T., Hirano Y., Serbulea V., Ambati M., Ambati V.L., Kajiwara Y., Ambati K., Bastos-Carvalho A., Ogura Y., Terasaki H., Oshika T., Kim K.B., Hinton D.R., Leitinger N., Cambier J.C., Buxbaum J.D., Kenney M.C., Jazwinski S.M., Nagai H., Hara I., West A.P., Fitzgerald K.A., Sadda S.R., Gelfand B.D, & Ambati J. (2017). cGAS drives non-canonical inflammasome activation in age-related macular degeneration. Nature medicine, 24(1), 50-61.
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4

Inflammasome Activation in Macrophages

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Primary peritoneal macrophages from C57BL/6 mice or THP1 cells were seeded in 24-well (5 × 105) or 6-well (2 × 106) culture plates. The next day, we removed medium and primed macrophages with LPS (InvivoGen tlrl-3pelps,100 ng/ml) for 3 h followed by 1,2-diol for 1 h. At the last, we treated macrophages with stimulation as follows: ATP (InvivoGen tlrl-atp, 5 mmol/L) or nigericin (InvivoGen tlrl-nig, 10 μmol/L) for 1 h and MSU (InvivoGen tlrl-msu, 200μg/ml) or SiO2 (InvivoGen tlrl-sio, 20 μg/ml) for 6 h. THP-1 cells should be treated with 100 ng/ml PMA for 12 h before seeding in culture plates.
Li J., Bai Y., Tang Y., Wang X., Cavagnaro M.J., Li L., Li Z., Zhang Y, & Shi J. (2022). A 4-Benzene-Indol Derivative Alleviates LPS-Induced Acute Lung Injury Through Inhibiting the NLRP3 Inflammasome. Frontiers in Immunology, 13, 812164.
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5

THP-1 Macrophage Activation Protocol

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For experiments, THP-1 cells were seeded at a density of 1 × 106 cells/ml and differentiated into macrophages using 100 μg/ml PMA (Sigma-Aldrich, P1585) for 48 h. Medium was changed to serum-free medium 6 h before transfection with MDP (100 ng/ml; InvivoGen, tlrl-mdpc), or to serum-free medium containing ultrapure lipopolysaccharide (upLPS, 100 ng/ml; InvivoGen, tlrl-3pelps) 16 h before treatment with MSU (150 μg/ml; InvivoGen, tlrl-msu) or ATP (2 mM; InvivoGen tlrl-atp). For transfection, MDP (InvivoGen, tlrl-mdpc) was dissolved in DMSO and mixed with Lipofectamine® (InvivoGen, L3000–001) for 15 min before applying to the cell culture media. As a control, DSMO mixed with Lipofectamine® was used. MSU and ATP were applied onto the cells as suspensions. Likewise, BMDCs were collected at d 7 of BM cultures in differentiation medium, and seeded in new plates (0.5 × 106 cells/well in 12-well plates) in RPMI supplemented with 10% FCS. After 6 h, medium was changed for serum-free medium 6 h before transfection with MDP or treatment with MSU or ATP as described above.
Spalinger M.R., Lang S., Gottier C., Dai X., Rawlings D.J., Chan A.C., Rogler G, & Scharl M. (2017). PTPN22 regulates NLRP3-mediated IL1B secretion in an autophagy-dependent manner. Autophagy, 13(9), 1590-1601.
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