Escherichia coli o111 b4

Manufactured by Merck Group

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Escherichia coli O111:B4 is a strain of the bacterium Escherichia coli. It is a bacterial endotoxin commonly used in laboratory research and testing.

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Lab products found in correlation

Lipopolysaccharides (lps), by Merck Group (11 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions) Opti mem, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Gemini Bio (3 mentions) Dmem f12, by Thermo Fisher Scientific (3 mentions) Co2 incubator, by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Griess reagent, by Merck Group (2 mentions) Periodate, by Merck Group (2 mentions) Periodate, by Solarbio (2 mentions) Sodium acetate, by Merck Group (2 mentions) Lps extraction kit, by iNtRON Biotechnology (2 mentions) Proteinase k, by Solarbio (2 mentions) Ifn γ, by Thermo Fisher Scientific (2 mentions) Endospecy es 50m kit, by Seikagaku (2 mentions) Tnf α elisa kit, by R&D Systems (2 mentions) Vacutainer, by BD (2 mentions) Dulbecco phosphate buffered saline (dpbs), by Merck Group (2 mentions) M csf, by BioLegend (2 mentions)

Close spelling variants of this product

LPS from Escherichia coli O111:B4 (161 citations) Escherichia coli (94 citations) LPS (Escherichia coli O111:B4, (91 citations) Lipopolysaccharide (LPS) from Escherichia coli O111:B4 (37 citations) Lipopolysaccharides (LPS) from Escherichia coli O111:B4 (23 citations) Lipopolysaccharides from Escherichia coli O111:B4 (20 citations) LPS from Escherichia coli serotype O111:B4 (16 citations) LPS (Escherichia coli serotype O111:B4; (14 citations) Lipopolysaccharides (LPS; Escherichia coli O111:B4) (5 citations) LPS purified from Escherichia coli O111:B4 (5 citations)

130 protocols using escherichia coli o111 b4

Sepsis-Induced Immune Paralysis Evaluation

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PBMCs and splenocytes were isolated at each time point after sepsis induction. PBMCs were isolated using the Ficoll gradient method [27 (link)]. Isolated PBMCs were stimulated with LPS to observe and compare the levels of immune paralysis. Tumor necrosis factor (TNF)-α levels were measured 5 h after LPS stimulation. Isolated PBMCs were seeded at a density of 1 × 10⁵ cells/mL in 96-well plates, and 100 ng/mL LPS (Escherichia coli O111: B4, Sigma-Aldrich, St. Louis, MO, USA) was added to each well. After 5 h, the culture medium was collected, and TNF-α levels were analyzed using a TNF-α enzyme-linked immunosorbent assay (ELISA) kit (ab236712, Abcam, Cambridge, MA, USA).
Splenocytes were isolated and stimulated with LPS to compare immune paralysis [28 (link)]. TNF-α levels were measured 5 h after LPS stimulation. Isolated splenocytes were seeded at a density of 5 × 10⁵ cells/mL in 6-well plates, and 1 μg/mL LPS (Escherichia coli O111: B4, Sigma-Aldrich, St. Louis, MO, USA) was added to each well. After 5 h, the culture medium was collected, and TNF-α was analyzed using a TNF-α ELISA kit (R&D Systems, Inc., Minneapolis, MN, USA).

Lee M.J., Bae J., Lee J.H., Park Y.J., Lee H.A., Mun S., Kim Y.S., Yune C.J., Chung T.N, & Kim K. (2022). Serial Change of Endotoxin Tolerance in a Polymicrobial Sepsis Model. International Journal of Molecular Sciences, 23(12), 6581.

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LPS-Induced Lung Injury Mouse Model

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LPS challenge was performed as previously described (de Souza Xavier Costa et al., 2017 (link)). Briefly, mice were exposed to nebulized LPS (Escherichia coli O111:B4, purified by phenol extraction; Sigma-Aldrich) at 0.8 mg/ml diluted in NaCl 0.9% (B Braun Medical, Sempach, Switzerland) in a plexiglas chamber connected to a nebulizer (System Assistance Medicale, Ledat, France) for 30 min. Control mice inhaled NaCl 0.9% only. Vehicle or ACT-1004-1239 was given p.o., 1 h prior (preventive setting) or 3 h post inhalation (therapeutic setting).
At different time points indicated in the figure legends (5, 24, 48, 72 h) following LPS or NaCl challenge, mice were euthanized with an overdose (150 mg/kg, intraperitoneally) of pentobarbital (Esconarkon, Streuli Pharma SA, Uznach, Switzerland), and samples were collected.

Pouzol L., Sassi A., Baumlin N., Tunis M., Strasser D.S., Lehembre F, & Martinic M.M. (2021). CXCR7 Antagonism Reduces Acute Lung Injury Pathogenesis. Frontiers in Pharmacology, 12, 748740.

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Differentiation of Monocytes into Macrophages

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Human peripheral blood CD14+ monocytes were purchased from LONZA (Lonza Walkersville, MD). Upon arrival, aliquots were thawed at 37°C and transferred to 50 mL conical tubes. Cells were washed twice with 10 mL of supplemented RPMI (10% FBS, 1% penicillin/streptomycin) and 20 U/mL of DNase I (Sigma Aldrich, St. Louis, MO), followed by centrifugation at 200 × g for 15 min. Cells were then re-suspended in RPMI containing 10% FBS, 1% penicillin/streptomycin, 1% sodium pyruvate, and 100 ng/mL of recombinant human M-CSF (eBioscience, San Diego, CA). M-CSF was used to differentiate monocytes into macrophages. Primary cells were cultured for 5–6 days in a 75-cm² tissue culture flask at 37°C with 5% of CO₂. Macrophages were harvested, and non-adherent cells and media were removed. The cells remaining in the culture flask were detached by adding 12 mL of trypsin for 30 min (37 °C, 5% CO₂). Cells were plated at 250,000 cells/mL in 24-well plates, seeded for 1 hour, stimulated with LPS (5 μg/mL, Escherichia coli O111:B4, Sigma), and transfected with a plasmid containing the CD163 gene or an empty vector. Supernatants and cells were harvested at 48 and 96 hours after LPS stimulation and stored at −80°C until used.

Alvarado-Vazquez P.A., Bernal L., Paige C.A., Grosick R.L., Vilrriales C.M., Ferreira D.W., Ulecia-Morón C, & Romero-Sandoval E.A. (2017). Macrophage-specific nanotechnology driven CD163 overexpression in human macrophages results in an M2 phenotype under inflammatory conditions. Immunobiology, 222(8-9), 900-912.

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LPS-induced IFN-β production in Fos knockout mice

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Fos knockout (FosKO) mice and LysM-Cre controls, aged between 10 and 14 wk, were injected i.p. with 5 mg/kg LPS (Escherichia coli O111:B4, Sigma-Aldrich). After 4 h, mice were sacrificed via cervical dislocation and blood was extracted from the heart using a 25G needle and 0.3-ml syringe. Blood was left to coagulate for 30 min and then centrifuged at 17,000 × g for 1 min. Serum was stored at −80°C until the assay for IFN-β by ELISA.

Blair L., Pattison M.J., Chakravarty P., Papoutsopoulou S., Bakiri L., Wagner E.F., Smale S, & Ley S.C. (2022). TPL-2 Inhibits IFN-β Expression via an ERK1/2-TCF-FOS Axis in TLR4-Stimulated Macrophages. The Journal of Immunology Author Choice, 208(4), 941-954.

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Bovine Endotoxin Challenge Model

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On d 111 postpartum, cows were challenged with standardized LPS by intravenous injection of 0.5 µg E. coli LPS per kg body weight (Escherichia coli O111:B4, Sigma Aldrich, St. Louis, Missouri, USA). Animals were monitored continuously after LPS administration and clinical observations were published previously [20 (link)].

Xu W., Grindler S., Kenéz Á., Dänicke S., Frahm J, & Huber K. (2022). Changes of the liver metabolome following an intravenous lipopolysaccharide injection in Holstein cows supplemented with dietary carnitine. Journal of Animal Science and Biotechnology, 13, 94.

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Investigating MAC-T Cell Responses to LPS and DAE

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The MAC-T cells used in this study were kindly provided by Dr. Jianxin Liu and Dr. Hongyun Liu at the College of Animal Science, Zhejiang University, China. Cells were cultured in a CO₂ incubator (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C, and culture medium was comprised of 90 mL of DMEM/F12 medium (Gibco, Grand Island, NY, USA), 10 mL of fetal bovine serum (Gibico, Grand Island, NY, USA), and 1 mL of penicillin-streptomycin solution (Hyclone, Logan, UT, USA).
MAC-T cells were firstly seeded into the culture medium without LPS and DAE, and after growing to 60% confluence the cells were divided into six groups or treatments (n = 6 per treatment) including the control, LPS (100 ng/mL LPS, Sigma, Escherichia coli O111:B4, Saint Louis, MO, USA), DAE10 (100 ng/mL LPS and 10 μg/mL DAE), DAE50 (100 ng/mL LPS and 50 μg/mL DAE), DAE100 (100 ng/mL LPS and 100 μg/mL DAE), and DAE200 (100 ng/mL LPS and 200 μg/mL DAE). MAC-T cells in each group were treated for 48 h, and culture medium was replaced every 24 h. The cells were collected after the treatment for further analysis.

Sun Y., Wu Y., Wang Z., Chen J., Yang Y, & Dong G. (2020). Dandelion Extract Alleviated Lipopolysaccharide-Induced Oxidative Stress through the Nrf2 Pathway in Bovine Mammary Epithelial Cells. Toxins, 12(8), 496.

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NEC Induction in C57BL/6J Mice

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All protocols and animal experiments were approved by the Indiana University Institutional Animal Care and Use Committee (IACUC). The NEC model used was extrapolated from a rat animal as described by Zani et al [18 (link)]. C57BL/6J mice were used for this experiment. Three groups were studied: (1) breastfed controls (n=11), formula-fed controls (n=15), and NEC (n=13). The total n in each group is made up of multiple litters, so each group was repeated at least once. Breastfed controls remained with dams and breastfed ad lib from postnatal day 5 (P5) until postnatal day 9 (P9). Formula-fed controls were taken from dams on P5 and given 300 kcal/kg daily of Esbilac milk replacer (PetAg, Hampshire, IL) fortified with Similac advance powder (Similac, Columbus, OH). Pups were fed via gavage three times per day with a 1.9 French catheter until P9. Pups undergoing the NEC experiment were taken from dams on P5. Experimental NEC was induced by administering 300 kcal/kg formula daily with 8 mg/kg lipopolysaccharide (LPS) isolated from Escherichia Coli O111:B4 (Cat # L4391, Sigma-Aldrich Company). Pups also received stress via hypothermia at 4° C for 12 minutes twice daily, and hypoxia at 5% O₂ for 10 minutes three times daily.

Hosfield B., Drucker N., Pecoraro A., Shelley W., Li H., Baxter N., Hawkins T, & Markel T.A. (2021). The Assessment of Microbiome Changes and Fecal Volatile Organic Compounds during Experimental Necrotizing Enterocolitis. Journal of pediatric surgery, 56(6), 1220-1225.

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HT Pretreatment Modulates LPS-Induced Response

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Cell were pretreated with HT (10 μM and 25 μM) (Sigma-Aldrich, Milano, Italy). One hour after HT pretreatment, cells were stimulated with LPS 1 μg/mL (Escherichia coli O111:B4, Sigma-Aldrich, Milano, Italy) for 1 or 6 h as previously described [47 (link)]. LPS concentrations were chosen based on previous studies by others using mammary epithelial cells [48 (link),49 (link)].

Fusco R., Cordaro M., Siracusa R., Peritore A.F., D’Amico R., Licata P., Crupi R, & Gugliandolo E. (2020). Effects of Hydroxytyrosol against Lipopolysaccharide-Induced Inflammation and Oxidative Stress in Bovine Mammary Epithelial Cells: A Natural Therapeutic Tool for Bovine Mastitis. Antioxidants, 9(8), 693.

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Evaluating Immunogenicity of Viral-Like Particles

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The immunogenic potential of VLPs was estimated by using RAW-Blue cells (Invivogen, San Diego, CA). RAW-Blue cells are derived from murine macrophage cell line RAW 264.7. These cells express a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of NF-κB and AP-1 promoters, two transcription factors that play en central role in inflammation and immunity. RAW-Blue cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) and 100 µg/mL Normocin. The cells (1 × 10⁵) were incubated with 25 μg/mL of P22CYP-PpIX-PEG(EST) or P22CYP preparations for 12 h at 37 °C and 5% CO₂. Supernatants were collected and SEAP production was evaluated based on the activity of alkaline phosphatase (AP) after the addition of 150 µL of quantity blue and the absorbance was determined at 655 nm. As positive control, 2.5 µg of purified lipopolysaccharides from Escherichia coli O111:B4 (Sigma-Aldrich) was used. The experiments were carried by triplicate.

Chauhan K., Hernandez-Meza J.M., Rodríguez-Hernández A.G., Juarez-Moreno K., Sengar P, & Vazquez-Duhalt R. (2018). Multifunctionalized biocatalytic P22 nanoreactor for combinatory treatment of ER+ breast cancer. Journal of Nanobiotechnology, 16, 17.

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Neuroinflammation in 5XFAD Mouse Model

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5XFAD and control C57BL/6J mice were maintained at Vanderbilt University under standard conditions, in a 12-h light/dark cycle, and with free access to food and water. The 5XFAD mice over express both mutant human amyloid precursor protein (APP) and presenilin 1 (PS1), correlating with high burden and accelerated accumulation of the Aβ. A colony of 5XFAD transgenic mice obtained from Jackson Laboratories was maintained by crossing 5XFAD mice with a WT C57BL/6J strain. The 5XFAD mice were maintained as heterozygous.
The mouse model of LPS-induced neuroinflammation was developed based on past reports^{50 (link),51 (link)}. The LPS derived from Escherichia coli O111:B4 (Sigma Aldrich, St Louis, MO) was formulated in sterilized dd. water and given a high dose of LPS formulation (5 mg/kg) through intraperitoneal injection 24 h before PET imaging. This high dose would result in approximately 12–13% body weight loss over the course of 24 h. After PET imaging, animals were sacrificed, and the brains were collected for histology analysis.

Behof W.J., Whitmore C.A., Haynes J.R., Rosenberg A.J., Tantawy M.N., Peterson T.E., Harrison F.E., Beelman R.B, & Pham W. (2021). A novel antioxidant ergothioneine PET radioligand for in vivo imaging applications. Scientific Reports, 11, 18450.

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