Lps e coli o111 b4

Manufactured by Merck Group

Sourced in United States, United Kingdom, Germany

LPS (E. coli O111:B4) is a laboratory reagent produced by Merck Group. It is a lipopolysaccharide extracted from the cell wall of Escherichia coli O111:B4 bacteria. This product is commonly used in research applications that require a source of endotoxin for in vitro studies.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Ifn γ, by R&D Systems (2 mentions) Nos2 mice, by Jackson ImmunoResearch (2 mentions) C57bl 6j mice, by Jackson ImmunoResearch (2 mentions) Ifn γ, by Abcam (2 mentions) M csf, by Thermo Fisher Scientific (2 mentions) Plp peptide, by Bio Basic (2 mentions) Mycobacterium tuberculosis h37ra, by BD (2 mentions) L4391 1mg, by Merck Group (2 mentions) P7208 50ug, by Merck Group (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) Pam3csk4, by InvivoGen (2 mentions) Nano itc, by Waters Corporation (2 mentions) Recombinant mouse ifn γ, by R&D Systems (2 mentions) Duoset elisa kit, by R&D Systems (2 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

E. coli LPS (149 citations) E. coli O111:B4 (78 citations) LPS O111:B4 (55 citations) Lipopolysaccharides (LPSs) from E. coli O111:B4 (2 citations)

94 protocols using lps e coli o111 b4

Alleviation of LPS-Induced Sepsis in Mice

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Female BALB/c mice (5-6 weeks) was provided by HFK Bioscience Co. (Beijing, China). All mice were raised in laminar flow cabinets and housed individually in a room with strictly controlled temperature and humidity, where a 12-hour light and 12-hour dark cycle was implemented, and mice had free access to food and water throughout the experimental period.
E. coli O111:B4 LPS (Sigma-Aldrich) at a concentration of 15 mg/L was injected into the peritoneal cavity of female BALB/c mice (age 7 weeks, weight 15 ± 0.5 g), and approximately 30 min after the challenge of E. coli O111:B4 LPS, MccJ25 was orally gavaged to the mice at a concentration of 4.55 or 9.1 mg per kg body weight (BW). The concentrations of MccJ25 were selected based on our previous study (24 (link)). Mice were followed for 72 h to test the survival rate of mice that were later sacrificed. However, for LPS treated mice, when the lifespan of the mice was around 50%, the mice were sacrificed. The serum from mice were obtained. The lung, liver, spleen, jejunum, and ileum were harvested, kept in liquid nitrogen immediately, stored at -80°C until analysis. For histopathological assessment, tissues and organs were fixed using 4% paraformaldehyde, hematoxylin eosin (H&E) was applied to examine the histological and pathological changes according to the standard protocol as described previously (24 (link)).

Yu H., Shang L., Yang G., Dai Z., Zeng X, & Qiao S. (2022). Biosynthetic Microcin J25 Exerts Strong Antibacterial, Anti-Inflammatory Activities, Low Cytotoxicity Without Increasing Drug-Resistance to Bacteria Target. Frontiers in Immunology, 13, 811378.

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Simulating Innate Immune Responses

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To simulate infection, the monolayers were challenged with 10 µg/mL of synthetic analog of dsRNA (poly I:C) (Sigma Aldrich, St. Louis, MO, USA), 50 ng/mL LPS (E. coli O111:B4, Merck, Rahway, NJ, USA), and 50 ng/mL Pam3Cys (Sigma Aldrich, St. Louis, MO, USA), respectively. Poly (I:C) was introduced into the cells using Lipofectamine 3000 (Thermo Fisher Scientific, Waltham, MA, USA). The cells were harvested four hours post challenge for RNA isolation.

Knez Š., Narat M, & Ogorevc J. (2022). Differential Gene Expression Induced by Different TLR Agonists in A549 Lung Epithelial Cells Is Modulated by CRISPR Activation of TLR10. Biomolecules, 13(1), 19.

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Profiling Oxidative Stress Markers

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8-iso-PGF_2α, PGF_2α, 8-iso-PGF_2α-d₄, and PGF_2α-d₄ were purchased from Cayman (Ann Arbor, MI, USA). Indomethacin, meclofenamic acid, carbon tetrachloride, m ethanol, ethanol, and acetic acid were purchased from Sigma Aldrich (St. Louis, MO, USA). LPS (E. coli, O111:B4) was purchased from EMD Millipore (Billerica, MA, USA).

van ‘t Erve T.J., Lih F.B., Jelsema C., Deterding L.J., Eling T.E., Mason R.P, & Kadiiska M.B. (2016). Reinterpreting the best biomarker of oxidative stress:The 8-iso-prostaglandin F2α/prostaglandin F2α ratio shows complex origins of lipid peroxidation biomarkers in animal models. Free radical biology & medicine, 95, 65-73.

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LPS Stimulation of Cell Cultures

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LPS (E. coli O111:B4) was purchased from EMD Millipore Co. (Billerica, MA, USA). A total of 5 mg of LPS was dissolved in 2 mL of PBS (-) and stored at −80 °C in the dark until use. After the treatment, LPS solution (1 ng/mL) was added to each well and incubated for 16 h at 37 °C.

Takahashi S., Ferdousi F., Yamamoto S., Hirano A., Nukaga S., Nozaki H, & Isoda H. (2023). Botryococcus terribilis Ethanol Extract Exerts Anti-inflammatory Effects on Murine RAW264 Cells. International Journal of Molecular Sciences, 24(7), 6666.

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Intestinal Inflammation Modulation by Peptides

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Seventy-two C57/BL6 male mice (6–8 weeks of age) were purchased from Charles River (Beijing, China). Mice were maintained in a specific pathogen free (SPF) environment at 22 ± 1°C with relative 55 ± 10% humidity, and the assays were performed in conformity with the laws and regulations for live animal treatment at China Agricultural University.
The mice were randomly distributed into six groups (n = 12 each): control, LPS (E. coli, O111:B4, Sigma-Aldrich, USA) treatment, LL-37 pretreatment followed by LPS treatment (LL-37 + LPS), Tα1 pretreatment followed by LPS treatment (Tα1 + LPS), LTA pretreatment followed by LPS treatment (LTA + LPS). Different peptides (10 mg/kg mouse weight) were injected intraperitoneally once daily for 7 days, whereas an equal volume of sterile saline was injected intraperitoneally to the control and LPS-treated groups. On day 7, mice in LPS, LL-37 + LPS, Tα1 + LPS, and LTA + LPS groups were intraperitoneally injected with LPS (10 mg/kg mouse weight) 1h after saline or the peptides treatment, and the control group was intraperitoneally injected with an equal volume of saline. The mice were then euthanized by cervical dislocation 6 h after intraperitoneal injection of LPS or saline, and samples of the intestines were collected for analysis. The body weights of the mice were recorded before and after the experiment.

Zhang L., Wei X., Zhang R., Petitte J.N., Si D., Li Z., Cheng J, & Du M. (2019). Design and Development of a Novel Peptide for Treating Intestinal Inflammation. Frontiers in Immunology, 10, 1841.

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Human and Rat Cell Culture Protocols

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Human astrocyte 1321N1 cells and rat neuron PC12 cells were purchased from the European Collection of Authenticated Cell Cultures (ECACC, Porton Down, UK). 1321N1 cells were cultured in Dulbecco’s modified Eagle’s Medium (DMEM) (Gibco; Invitrogen, USA) supplemented with 10% fetal bovine serum and 100 U/ml penicillin-streptomycin. Some cohort 1321N1 cultures were treated with lipopolysaccharide (LPS) E. coli O111:B4 (100 ng/ml, Sigma-Aldrich, Poole, UK) for 24 h, some cohort cultures were treated with 1 µM dexmedetomidine (Sigma-Aldrich) 30 min before LPS treatment^{22 (link)}, and some cohort cultures were treated with TNF-α (100 ng/ml, Sigma-Aldrich) for 24 h. PC12 cells were maintained in RPMI-1640 medium (Gibco; Invitrogen, USA) containing 10% heat-inactivated horse serum, 5% fetal bovine serum and 100 U/ml penicillin-streptomycin. To induce the differentiation of PC12 cells into neuronal like cells, 50 ng/ml nerve growth factor (NGF) was added into the medium with 1% horse serum and cells were seeded on a Collagen IV coated dish for 24 h. The medium was routinely replaced with fresh medium every 2 days for 5 days until neuronal differentiation had taken place. PC12 cells were treated with calf thymus histones (1000 ng/ml, Sigma-Aldrich) for 24 h. The cells were incubated at 37 °C in a humidified atmosphere with 5% CO₂.

Sun Y.B., Zhao H., Mu D.L., Zhang W., Cui J., Wu L., Alam A., Wang D.X, & Ma D. (2019). Dexmedetomidine inhibits astrocyte pyroptosis and subsequently protects the brain in in vitro and in vivo models of sepsis. Cell Death & Disease, 10(3), 167.

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Differentiation and LPS Treatment of PC12 Cells

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Rat pheochromocytoma PC12 cells were purchased from the China Infrastructure of Cell Line Resource (Beijing, China) and cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco; Invitrogen, Waltham, MA, USA) supplemented with 10% horse serum (Invitrogen), 5% fetal bovine serum (Invitrogen), and 100 μg/ml penicillin/streptomycin (Invitrogen) at 37°C with 5% CO₂. The cells were maintained in culture dishes precoated with 0.5 mg/ml poly-D-lysine (Sigma, St. Louis, MO, USA) to improve cell adherence. The medium was replaced by DMEM containing 1% horse serum and 50 ng/ml nerve growth factor (NGF) (Sigma) to induce neural cell differentiation 24 h after seeding. Half of the NGF medium was refreshed every other day. Five days later, the differentiated PC12 cells (Figure S1b) were treated with LPS E. coli O111:B4 (1 μg/ml; Sigma).

Li Y., Yuan Y., Li Y., Han D., Liu T., Yang N., Mi X., Hong J., Liu K., Song Y., He J., Zhou Y., Han Y., Shi C., Yu S., Zou P., Guo X, & Li Z. (2021). Inhibition of α-Synuclein Accumulation Improves Neuronal Apoptosis and Delayed Postoperative Cognitive Recovery in Aged Mice. Oxidative Medicine and Cellular Longevity, 2021, 5572899.

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LPS and TAK 242 Preparation Protocol

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LPS (E. coli O111:B4) was purchased from Sigma-Aldrich (catalog No. L2630-25MG). This was dissolved in normal saline to make a stock solution of 5 mg/ml from which an aliquot was taken and diluted to a working solution of 0.25 mg/ml and 0.125 mg/ml. TAK 242 was purchased from Tocris bioscience (Catalog No. 6587). TAK 242 was first dissolved in ethanol and then further diluted with normal saline to a final concentration of 0.25 mg/ml, with an ethanol concentration of 2.5%.

Kisipan M.L., Ojoo R.O., Kanui T.I, & Abelson K.S. (2022). Bodyweight, locomotion, and behavioral responses of the naked mole rat (Heterocephalus glaber) to lipopolysaccharide administration. Journal of Comparative Physiology. A, Neuroethology, Sensory, Neural, and Behavioral Physiology, 208(4), 493-504.

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In vitro microglial cell study

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For in vitro studies, we used a microglial cell line SIM-A9 (T0247-GVO-ABM, BioCat, Heidelberg, Germany). Mouse microglial cells were cultured in 24-well microplates at a density of 1 × 105 cells per cm² in complete DMEM/F12 medium and incubated for 1 h at 37 °C with 5% CO₂. After cell adhesion, the culture medium was replaced with a medium containing the test DS (composition of N-acylethanolamines) in 0.1, 1, and 10 μg/mL concentrations and incubated at 37 °C with 5% CO₂ for one hour. Next, the solution of bacterial lipopolysaccharides (LPS, E. coli O111:B4, Sigma-Aldrich, Bellefonte, PA, USA) was added to the samples to obtain a concentration of 1 μg/mL and incubated at 37 °C with 5% CO₂ for 24 h. The experiment used a negative control (cells incubated in a normal culture medium poor DS) and control of LPS activity (cells incubated with LPS).

Tyrtyshnaia A., Konovalova S., Ponomarenko A., Egoraeva A, & Manzhulo I. (2022). Fatty Acid-Derived N-acylethanolamines Dietary Supplementation Attenuates Neuroinflammation and Cognitive Impairment in LPS Murine Model. Nutrients, 14(18), 3879.

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Inflammatory Pathway Activation Assays

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InVivoMab anti-mouse Ly6G (CAT#: BE0075-1) was purchased from Bio X Cell (Lebanon, NH, USA). Compound C (CpC, CAT#: 171260), LPS (E. coli O111:B4, CAT#: L4391), STO-609 (STO, CAT#: 570250) and lucigenin (CAT#: M8010) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse interleukin-1β (IL-1β)/IL-1F2 Quantikine ELISA Kit (CAT#: MLB00C) and Mouse IL-6 Quantikine ELISA Kit (CAT#: M6000B) were purchased from R&D Systems, Inc. (Minneapolis, MN, Canada). MojoSort™ Human Pan Monocyte Isolation Kit (CAT#: 480060) was purchased from BioLegend (USA). Human Macrophage Colony-Stimulating Factor (M-CSF) Recombinant Protein (CAT#: RP-8643), Pierce™ BCA Protein Assay Kit (CAT#: 23227) and Amplex Red Hydrogen Peroxide⁄Peroxidase Assay Kit (CAT#: A22188) were purchased from Invitrogen (Carlsbad, CA, USA). Lactate Dehydrogenase (LDH) Assay Kit (CAT#: A020-2), Myeloperoxidase (MPO) Assay Kit (CAT#: A044-1-1), ROS Assay Kit (CAT#: E004-1-1), Malondialdehyde (MDA) Assay Kit (CAT#: A003-1-1), 4-Hydroxynonenal ELISA Kit (CAT#: H268), Protein Carbonyl Assay Kit (CAT#: A087-1-1), Total Anti-oxidant Capacity Assay Kit (CAT#: A015-2-1) and Superoxide Dismutase (SOD) Assay Kit (CAT#: A001-3-2) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TransAM® NF-κB p65 Kit was purchased from Active Motif (Carlsbad, CA, USA).

Chen R., Cao C., Liu H., Jiang W., Pan R., He H., Ding K, & Meng Q. (2022). Macrophage Sprouty4 deficiency diminishes sepsis-induced acute lung injury in mice. Redox Biology, 58, 102513.

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