Lps e coli serotype r515

Manufactured by Enzo Life Sciences

LPS E.coli serotype R515 is a lipopolysaccharide extracted from Escherichia coli bacteria. It is commonly used as a toll-like receptor 4 (TLR4) agonist in cell-based assays and research studies.

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Lab products found in correlation

2 deoxyglucose, by Merck Group (1 mentions) Cp 91149, by Merck Group (1 mentions) Oligomycin a, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) C57bl 6 mice, by Charles River Laboratories (1 mentions) Thapsigargin, by Bio-Techne (1 mentions) Dorsomorphin, by Bio-Techne (1 mentions) Gsk2606414, by Bio-Techne (1 mentions) Gsk2578215a, by Bio-Techne (1 mentions) Anti mouse cd38, by BioLegend (1 mentions) Bapta am, by R&D Systems (1 mentions) Egta am, by AnaSpec (1 mentions) Rtk2758, by BioLegend (1 mentions) Naadp am, by AAT Bioquest (1 mentions) 8 br adpr, by Biolog (1 mentions) Chir99021, by Bio-Techne (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)

3 protocols using lps e coli serotype r515

Neutrophil Metabolic Reprogramming Assay

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Neutrophils were cultured at 5 million cells/mL for 2-20 h at 37°C, in normoxia (21% O2, 5% CO2) or hypoxia (1% (3kPa) O2, 5% CO2) in two types of pre-equilibrated RPMI media 1640 with 10% fetal bovine serum (FBS) (GIBCO) and 1% Penicillin/Streptomycin, differing in the presence or absence of glucose. In the absence of glucose, dialyzed glucose-free FBS was used. To study the effects of stimulation and metabolic inhibitors, neutrophils were cultured with (100ng/mL) LPS E.coli serotype R515 (Enzo), CP-91149 (Sigma-Aldrich), 2-deoxyglucose (Sigma-Aldrich), Oligomycin A (Sigma-Aldrich) and Etomoxir (10uM) (CNIO Carlos III Therapies) (Raud et al., 2018 (link); Yao et al., 2018 (link)), Fructose-1,6-bisphosphatase-1 Inhibitor (FBP1i) (Cambridge Bioscience), MB05032 (Tocris Bioscience), 3-Mercaptopicolinic acid (HCl) (3-MPA) (Cambridge Bioscience), Glutaminase Inhibitor II, BPTES (MERCK Chemical).

Sadiku P., Willson J.A., Ryan E.M., Sammut D., Coelho P., Watts E.R., Grecian R., Young J.M., Bewley M., Arienti S., Mirchandani A.S., Sanchez Garcia M.A., Morrison T., Zhang A., Reyes L., Griessler T., Jheeta P., Paterson G.G., Graham C.J., Thomson J.P., Baillie K., Thompson A.A., Morgan J.M., Acosta-Sanchez A., Dardé V.M., Duran J., Guinovart J.J., Rodriguez-Blanco G., Von Kriegsheim A., Meehan R.R., Mazzone M., Dockrell D.H., Ghesquiere B., Carmeliet P., Whyte M.K, & Walmsley S.R. (2021). Neutrophils Fuel Effective Immune Responses through Gluconeogenesis and Glycogenesis. Cell Metabolism, 33(2), 411-423.e4.

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Murine Model of Inflammation

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C57BL/6 mice were purchased from Charles River at 6–8 weeks of age. Animals were housed in a licensed BioResource Unit (Dublin City University), according to Health Products Regulatory Authority (HPRA) regulations and had ad libitum access to animal chow and water. LPS E. coli serotype R515 was purchased from Enzo Life Sciences. Loxoribine and PGN from InvivoGen, rGMCSF from Sigma Aldrich, and STX3 siRNA from Life Technologies.

Collins L.E., DeCourcey J., Rochfort K.D., Kristek M, & Loscher C.E. (2015). A Role for Syntaxin 3 in the Secretion of IL-6 from Dendritic Cells Following Activation of Toll-Like Receptors. Frontiers in Immunology, 5, 691.

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Reagents for Immune Cell Stimulation

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The following antibodies were used for cell stimulation: purified anti-mouse CD38 (Clone 90; BioLegend, 102702), purified rat IgG2a, κ isotype control (Clone RTK2758; BioLegend, 400565). In certain experiments, the following reagents/inhibitors were used at the indicated concentration in cell culture: LPS (E. Coli Serotype R515; Enzo Life Sciences, ALX-581-007-L001), NAADP-AM (3 μM; AAT Bioquest, 21000), GSK2578215A (LRRK2 inhibitor, 1 µM; Tocris Bioscience, 4629), GPN (200 µM; Santa Cruz Biotechnology, sc-252858), EDTA (2 mM; Sigma-Aldrich, E6758), thapsigargin (500 nM; Tocris Bioscience, 1138), 8-Br-cADPR (100 µM; BIOLOG Life Science Institute, B065), 8-Br-ADPR (100 µM; BIOLOG Life Science Institute, B051), Ned-19 (100 µM; Cayman Chemical, 17527), cyclosporine (10 µM; R&D Systems, 1101/100), FK506 (5 nM; Eton Bioscience Inc, 1100060052), CHIR 99021 (GSK3B inhibitor, 5 µM; Tocris Bioscience, 4423), BAPTA-AM (10 µM; R&D Systems, 2787), EGTA-AM (10 µM; AnaSpec Inc, AS-84100), GSK2606414 (EIF2AK3/PERK inhibitor, 10 µM; Tocris Bioscience, 5107), and dorsomorphin (5 µM; Tocris Bioscience, 3093).

Nabar N.R., Heijjer C.N., Shi C.S., Hwang I.Y., Ganesan S., Karlsson M.C, & Kehrl J.H. (2021). LRRK2 is required for CD38-mediated NAADP-Ca2+ signaling and the downstream activation of TFEB (transcription factor EB) in immune cells. Autophagy, 18(1), 204-222.

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