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Lps from e coli serotype 0111 b4

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LPS (from E. coli serotype 0111:B4) is a laboratory reagent used in research applications. It is a component of the outer membrane of Gram-negative bacteria, and it is derived from Escherichia coli serotype 0111:B4. This product is intended for use in scientific research and experiments, and its core function is to serve as a standardized source of lipopolysaccharide for experimental purposes.

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Lab products found in correlation

Trypan blue dye, by Merck Group (3 mentions) Heat inactivated fetal calf serum, by Merck Group (3 mentions) L glutamine, by Merck Group (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Agarose, by Merck Group (2 mentions) High molecular weight poly 1 c, by InvivoGen (2 mentions) Aspirin, by Merck Group (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Hepes, by Merck Group (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Indomethacin, by Merck Group (1 mentions) Donepezil hydrochloride, by Merck Group (1 mentions) Assay kit for gsh, by Merck Group (1 mentions) Spermidine, by Merck Group (1 mentions) Gentisic acid, by Merck Group (1 mentions) Cm h2dcfda, by Thermo Fisher Scientific (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions) Propidium iodide, by Merck Group (1 mentions)

Spelling variants of this product

E. coli LPS (149 citations) LPS (E. coli 0111:B4) (106 citations) LPS from E. coli (104 citations) E. coli 0111:B4 (81 citations) LPS from E. coli 0111:B4 (52 citations) LPS (0111:B4, (31 citations) LPS (E. coli serotype 0111:B4; (15 citations) E. coli serotype 0111:B4 (15 citations) Lipopolysaccharide (LPS) derived from E. coli (serotype 0111:B4) (2 citations)

12 protocols using lps from e coli serotype 0111 b4

1

Generation of Murine Bone Marrow-Derived Macrophages

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Bone marrow cells were flushed from femurs and tibias of mice with 10 mL of DMEM supplemented with 10% FBS (Gibco), antibiotic/antimycotic solution (100 X, HyClone), GlutaMAX (100X, Gibco) and HEPES (25 mM, Sigma). Cells were pelleted by centrifugation and resuspended in ACK lysis buffer for 5 min and resuspended in DMEM. 107 cells were resuspended in DMEM supplemented with 20 ng/mL of M-CSF (Peprotech) and 5 ng/mL of GM-CSF (Peprotech) and plated in 10 cm tissue culture-treated petri dishes. Three days later, we added 10 mL of BMDM medium and on day 6, the medium was replaced with 10mL of fresh BMDM medium. 24 hours later, cells were treated with different doses of LPS from E. coli serotype 0111: B4 (Sigma-Aldrich) for different time points. In different experiments, cells were treated with anti-IL10R (10 μg, clone 1B1.3A BioXcel), cyclooxygenase inhibitors (Aspirin 100 μM, and Indomethacin 10 μM, Sigma Aldrich) and mPGES1 inhibitor CAY10526 (Cayman Chem.).
De Melo P., Rocio Pineros Alvarez A., Ye X., Blackman A., Alves-Filho J.C., Medeiros A.I., Rathmell J., Pua H, & Serezani C.H. (2021). Macrophage-derived microRNA-21 drives overwhelming glycolytic and inflammatory response during sepsis via repression of the PGE2/IL-10 axis.. Journal of immunology (Baltimore, Md. : 1950), 207(3), 902-912.
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2

Synthesis and Evaluation of H3R Antagonist DL77

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The H3R antagonist DL77 [1-(3-(4-tert-pentylphenoxy) propyl)piperidine] was designed and synthesized in the Department of Technology and Biotechnology of Drugs Krakow, Poland, according to a previously described procedure23 (link). Sodium valproate (VPA) (500 mg/kg, i.p.), donepezil hydrochloride (DOZ) (1 mg/kg, i.p.) (reference drug), the H3R agonist RAMH (10 mg/kg, i.p.), the brain-penetrant H1R antagonist PYR (10 mg/kg, i.p.), the brain-penetrant H2R antagonist ZOL (10 mg/kg, i.p.), LPS (from E. coli serotype 0111:B4), and the assay kit for GSH were all purchased from Sigma-Aldrich (St. Louis, MO, USA). The lipid peroxidation assay kit for estimation of malondialdehyde (MDA) was obtained from North West Life Science (Vancouver, WA, USA). For estimation of the proinflammatory cytokines IL-1β, IL-6 and TNF-α mediated by lipopolysaccharide (LPS), commercially available ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA). All the reagents used in the study were of analytical grade. All drugs were dissolved in isotonic saline solution and injected intraperitoneally (i.p.) at a volume of 10 ml/kg adjusted to body weight, and all doses are expressed in terms of the free base.
Eissa N., Jayaprakash P., Azimullah S., Ojha S.K., Al-Houqani M., Jalal F.Y., Łażewska D., Kieć-Kononowicz K, & Sadek B. (2018). The histamine H3R antagonist DL77 attenuates autistic behaviors in a prenatal valproic acid-induced mouse model of autism. Scientific Reports, 8, 13077.
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3

Antibiotics and Oxidative Stress Assays

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Antibiotics (ciprofloxacin, gentamicin, and ampicillin), 2,5-dihydroxybenzoic acid (gentisic acid), spermidine, HBTU, EDTA, DNTB, DAPI, propidium iodide (PI), carboxyfluorescein diacetate assay (CFDA), Triton X-100, LPS from E. coli serotype:0111:B4 and agarose were purchased from Sigma Chemical Co. (St. Louis, MO, United States), and CM-H2DCFDA was purchased from Thermo Fisher Scientific (Waltham, MA, United States).
Espinoza-Culupú A., Mendes E., Vitorino H.A., da Silva PI J.r, & Borges M.M. (2020). Mygalin: An Acylpolyamine With Bactericidal Activity. Frontiers in Microbiology, 10, 2928.
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4

Ethanol and LPS administration protocols

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When administered i.p., ethanol (95%) was diluted fresh daily with pyrogen-free physiological saline (0.9%, Teknova, Hollister, CA) to a final concentration of 20% (v/v) and sterile saline alone was used for vehicle. When delivered i.g., ethanol was mixed with tap water (20% v/v), with tap water alone delivered isovolumetrically to the ethanol intubations as the vehicle. LPS (from E. coli serotype 0111:B4, Sigma-Aldrich, St. Louis, MO) was initially diluted in sterile, pyrogen-free saline and aliquots were stored at −20 °C until needed. On the day of experimentation, a frozen aliquot was thawed and diluted to the required concentration [250 µg/kg (i.p. at 1.0 ml/kg)], also in pyrogen-free physiological saline.
Doremus-Fitzwater T.L., Gano A., Paniccia J.E, & Deak T. (2015). Male adolescent rats display blunted cytokine responses in the CNS after acute ethanol or lipopolysaccharide exposure. Physiology & behavior, 148, 131-144.
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5

Screening of TAS2R Agonists for Biological Activity

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The TAS2R agonists chloroquine diphosphate, quinine hydrochloride dihydrate, saccharin sodium hydrate, denatonium benzoate, 1,10-phenanthroline hydrochloride monohydrate, ofloxacin, strychnine hemisulphate, erythromycin, dapsone, carisoprodol, and sodium cromoglycate were obtained from Sigma-Aldrich (Saint-Quentin Fallavier, France) and diphenidol hydrochloride was provided by TCI Europe (Zwijndrecht, Belgium). All products were solubilized and diluted in sterile water, with the exception of erythromycin, dapsone, and carisoprodol, which were solubilized in DMSO and then diluted in water. Antibiotics, DMSO, L-glutamine, trypan blue dye, heat-inactivated fetal calf serum and LPS (from E. coli serotype 0111:B4) were purchased from Sigma (St. Louis, MO, United States). RPMI medium was from Eurobio Biotechnology (Les Ulis, France).
Grassin-Delyle S., Salvator H., Mantov N., Abrial C., Brollo M., Faisy C., Naline E., Couderc L.J, & Devillier P. (2019). Bitter Taste Receptors (TAS2Rs) in Human Lung Macrophages: Receptor Expression and Inhibitory Effects of TAS2R Agonists. Frontiers in Physiology, 10, 1267.
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6

Ruxolitinib and Budesonide Cytokine Assay

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Ruxolitinib was obtained from Selleck Chemicals LLC (Houston, TX), and budesonide was obtained from Sigma (St. Louis, MO). Both were dissolved in vehicle (0.05% dimethylsulfoxide), which did not interfere with cytokine production (data not shown). Antibiotics, dimethylsulfoxide, L-glutamine, trypan blue dye, heat-inactivated fetal calf serum, and LPS (from E. coli serotype 0111:B4) were purchased from Sigma. High-molecular-weight poly (I:C) was obtained from InvivoGen (Toulouse, France). Bovine serum albumin and Roswell Park Memorial Institute (RPMI) medium were purchased from Eurobio Biotechnology (Les Ulis, France).
Mantov N., Zrounba M., Brollo M., Grassin-Delyle S., Glorion M., David M., Naline E., Devillier P, & Salvator H. (2022). Ruxolitinib inhibits cytokine production by human lung macrophages without impairing phagocytic ability. Frontiers in Pharmacology, 13, 896167.
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7

Flow Cytometry Staining and Activation

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Fc receptors were blocked using anti-mouse CD16/32 (0.25 µg; eBioscience). Cells were then stained for 30 min at 4 °C using the specified antibodies (GM-CSF Rα from R&D Systems; all the others from BD Biosciences or eBioscience). To detect T cell cytokine expression, cells were activated for 18 h with 1 mg/ml ionomycin and 20 ng/ml phorbol 12-myristate 13-acetate 40 (PMA) in the presence of 2 mg/ml brefeldin A (Sigma) for the last 6 h of co-culture. To detect antigen-presenting cell cytokine expression, cells were activated for 18 h with 100 ng/ml LPS from E. coli serotype 0111:B4 (Sigma) in the presence of 2 mg/ml brefeldin A (Sigma) for the last 6 h of co-culture. Cells were stained for surface markers and a fixation and permeabilization kit (eBioscience) was used. Cells were acquired on a BD Canto II and analyzed using BD FACSDiva version 6.1 software.
Ifergan I., Davidson T.S., Kebir H., Xu D., Palacios-Macapagal D., Cann J., Rodgers J.M., Hunter Z.N., Pittet C.L., Beddow S., Jones C.A., Prat A., Sleeman M.A, & Miller S.D. (2017). Targeting the GM-CSF receptor for the treatment of CNS autoimmunity. Journal of autoimmunity, 84, 1-11.
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8

Antimicrobial Peptide Membrane Interactions

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DOPC and DOPG were from Avanti Polar Lipids (Alabaster, AL, USA). ANTS, DPX and BC were purchased from Invitrogen (Carlsbad, CA, USA). LPS from E. coli serotype 0111:B4 were purchased from Sigma-Aldrich (St. Louis, MO, USA). PD-10 desalting columns with Sephadex G-25 were from GE Healthcare (Waukesha, WI, USA). RNase6(1–45) peptide was purchased from Genecust (Dudelange, Luxembourg). Strains used were Escherichia coli (BL21; Novagen, Madison, WI, USA), Staphylococcus aureus (ATCC 502A; Manassas, VA, USA), Acinetobacter baumannii (ATCC 15308; Manassas, VA, USA), Pseudomonas aeruginosa (ATCC 47085; Manassas, VA, USA), Micrococcus luteus (ATCC 7468; Manassas, VA, USA) and Enterococcus faecium (ATCC 19434; Manassas, VA, USA).
Pulido D., Arranz-Trullén J., Prats-Ejarque G., Velázquez D., Torrent M., Moussaoui M, & Boix E. (2016). Insights into the Antimicrobial Mechanism of Action of Human RNase6: Structural Determinants for Bacterial Cell Agglutination and Membrane Permeation. International Journal of Molecular Sciences, 17(4), 552.
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9

Recombinant APN Production and Characterization

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Human recombinant APNs (expressed either in Escherichia coli (E.coli) or in human embryonic kidney (HEK) 293 cells) were purchased from Biovendor (Karasek, Czech Republic). AdipoRon hydrochloride and human recombinant IL-4 (produced in E.coli) were acquired from TOCRIS/R&D Systems (Lille, France) and solubilized respectively in DMSO and Roswell Park Memorial Institute 1,640 medium (RPMI). Antibiotics, DMSO, l-glutamine, trypan blue dye, heat-inactivated fetal calf serum and LPS (from E. coli serotype 0111:B4) were purchased from Sigma (St. Louis, MO, United States). High-molecular-weight poly (I:C) was obtained from InvivoGen (Toulouse, France). Bovine serum albumin and RPMI medium were from Eurobio Biotechnology (Les Ulis, France).
Salvator H., Grassin-Delyle S., Brollo M., Couderc L.J., Abrial C., Victoni T., Naline E, & Devillier P. (2021). Adiponectin Inhibits the Production of TNF-α, IL-6 and Chemokines by Human Lung Macrophages. Frontiers in Pharmacology, 12, 718929.
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10

Sepsis and Endotoxemia Induction Protocols

Cited 1 time
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Sepsis was induced by CLP as previously described (32 (link)). A 21 G needle was used to cause moderate sepsis. The survival rate was evaluated 6 h after CLP and twice per day every day during 7 days after CLP induction. Peritoneal lavage, blood, and lung were harvested 6 h after surgery to measure cytokines, markers of tissue damage, or bacterial burden in septic mice.
In another experimental setting, miR-21fl/fl and miR-21Δmyel mice were challenged i.p. with either a moderate (5mg/kg) or lethal dose (10 mg/kg) of LPS from E. coli serotype 0111: B4 (Sigma-Aldrich) or 0.9% saline control (33 (link)). Anti-IL10R treatment or isotype control (10 mg/kg, clone 1B1.3A BioXcel) was injected 1 hour before LPS injection (32 (link)). Mice survival was assessed over 7 days. Cytokine levels were measured in serum and/or peritoneal lavage fluid 12 hours after challenge with LPS.
De Melo P., Rocio Pineros Alvarez A., Ye X., Blackman A., Alves-Filho J.C., Medeiros A.I., Rathmell J., Pua H, & Serezani C.H. (2021). Macrophage-derived microRNA-21 drives overwhelming glycolytic and inflammatory response during sepsis via repression of the PGE2/IL-10 axis.. Journal of immunology (Baltimore, Md. : 1950), 207(3), 902-912.
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