Lps escherichia coli serotype 055 b5

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LPS (Escherichia coli serotype 055:B5) is a purified lipopolysaccharide extracted from the outer membrane of Escherichia coli bacteria. It is a common laboratory reagent used in biological research.

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LPS (Escherichia coli 055:B5) (171 citations) Escherichia coli LPS (168 citations) Escherichia coli (94 citations) Escherichia coli 055:B5 (72 citations) Escherichia coli serotype 055:B5 (36 citations) LPS (055: B5) (23 citations) LPS from Escherichia coli serotype 055:B5 (22 citations) Lipopolysaccharide (LPS) (Escherichia coli serotype 055:B5) (7 citations) LPS serotype 055:B5 (5 citations) Bacterial lipopolysaccharide (LPS) from Escherichia coli serotype 055:B5 (4 citations)

60 protocols using lps escherichia coli serotype 055 b5

Investigating Macrophage Inflammatory Responses

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Key reagents and their sources were as follows: Escherichia coli LPS serotype 055:B5, TNF-α protease inhibitor-0 (TAPI-0) and ATP were from Sigma-Aldrich; selective p38 inhibitor (SB202190) was from Calbiochem Merck-Millipore; P2X7 receptor selective antagonists AZ10606120, A438079, and A740003 were from Tocris. The composition of the physiological buffer used in all experiments to stimulate macrophages with ATP was (in millimoles): 147 NaCl, 10 HEPES, 13 d-glucose, 2 KCl, 2 CaCl₂, and 1 MgCl₂; pH 7.4.
HEK293T cells (ATCC CRL-11268) were cultured in DMEM:F-12 media (1:1; Lonza) supplemented with 10% of fetal calf serum (Life Technologies) and 2 mM Glutamax (Life Technologies) and were routinely tested for mycoplasma contamination with a Mycoplasma Detection Kit (Roche). Lipofectamine 2000 (Life Technologies) was used according to the manufacturer’s instructions to transfect a plasmid coding for human TNF-α into HEK293T cells.

Barberà-Cremades M., Gómez A.I., Baroja-Mazo A., Martínez-Alarcón L., Martínez C.M., de Torre-Minguela C, & Pelegrín P. (2017). P2X7 Receptor Induces Tumor Necrosis Factor-α Converting Enzyme Activation and Release to Boost TNF-α Production. Frontiers in Immunology, 8, 862.

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Steroid and Endotoxin Preparation Protocol

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Water‐soluble progesterone, β‐estradiol, 5α‐dihydrotestosterone (DHT) solution in methanol, hydrocortisone, mifepristone, PF‐02413873, spironolactone, and Escherichia coli LPS (serotype 055:B5) were all obtained from Sigma–Aldrich (St. Louis, MO, USA). LPS was further purified as described previously.¹³ Dexamethasone was obtained from Centrafarm (Breda, The Netherlands).

Groh L.A., Verel D.E., van der Heijden C.D., Matzaraki V., Moorlag S.J., de Bree L.C., Koeken V.A., Mourits V.P., Keating S.T., van Puffelen J.H., Joosten L.A., Netea M.G, & Riksen N.P. (2022). Immune modulatory effects of progesterone on oxLDL‐induced trained immunity in monocytes. Journal of Leukocyte Biology, 112(2), 279-288.

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Osteoblast Regulation via Redox Signaling

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MC3T3-E1 cells, an osteoblast-like cell line, was bought from the Shanghai Cell Center. Escherichia coli LPS (serotype 055:B5) and H2DCFDA were purchased from Sigma Chemical Co (St. Louis, MO, USA). NAC and NQDI-1 were purchased from MedChem Express (MCE, NJ, USA). Antibodies to Prx1, Trx, Bax, Bcl-2, JNK, P38, Cyto-c, p-JNK, P-P38, ASK1 and Caspase 3 were purchased from Abcam (Shanghai, China). Anti-GAPDH and anti-p-ASK1 were purchased from Proteintech (Wuhan, Hubei, China) and Affinity (OH, USA) respectively.

Feng H., Li Z., Du J., Sun J., Feng W., Li D., Liu S., Wang W., Liu H., Amizuka N, & Li M. (2018). Dual function of peroxiredoxin I in lipopolysaccharide-induced osteoblast apoptosis via reactive oxygen species and the apoptosis signal-regulating kinase 1 signaling pathway. Cell Death Discovery, 4, 47.

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Anti-inflammatory Agent Evaluation

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α-Bisabolol (anti-irritant and anti-inflammatory agent), forskolin (direct AC activator), Escherichia coli LPS (serotype 055:B5), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibiotics, penicillin-streptomycin (10,000 U/ml), Dulbecco's modified Eagle's medium and Dulbecco phosphate buffered saline (PBS 10×) were purchased in Mexico from Gibco, ThermoFisher Scientific.

Muñoz-Pérez V.M., Ortiz M.I., Ponce-Monter H.A., Monter-Pérez V, & Barragán-Ramírez G. (2018). Anti-inflammatory and utero-relaxant effect of α-bisabolol on the pregnant human uterus. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 22(4), 391-398.

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Rat LPS Injection Protocol

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The model was established as previously described [40 (link)–42 (link)]. The rats were injected with LPS (Escherichia coli, serotype 055: B5; Sigma–Aldrich, St Louis, MO, USA) (300 μg/kg, intraperitoneally) at gestational Days 11, 14 and 18.

Li Y., Luo W., Zhang J., Luo Y., Han W., Wang H., Xia H., Chen Z., Yang Y., Chen Q., Li H., Yang L., Hu C., Huang H., Peng Z., Tan X., Li M, & Yang J. (2022). Maternal Inflammation Exaggerates Offspring Susceptibility to Cerebral Ischemia–Reperfusion Injury via the COX-2/PGD2/DP2 Pathway Activation. Oxidative Medicine and Cellular Longevity, 2022, 1571705.

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β-Lapachone and LPS-Induced Inflammatory Response

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All reagents for cell culture were purchased from Gibco BRL (Grand Island, NY, USA). β-Lapachone and LPS (Escherichia coli serotype 055:B5) were obtained from Sigma–Aldrich (St. Louis, MO, USA). All reagents and enzymes for reverse transcription polymerase chain reaction (RT-PCR) and oligonucleotides for electrophoretic mobility shift assay (EMSA) were purchased from Promega (Madison, WI, USA). Antibodies against phospho-/total forms of MAPKs, CREB, β-actin, MMPs (MMP-3, MMP-8, MMP-9), and tissue inhibitor of metalloproteinase-2 (TIMP-2) were supplied by Cell Signaling Technology (Beverley, CA, USA), Abcam (Cambridge, UK), or Chemicon (Temecula, CA, USA). Antibodies against HO-1, NQO1, and Iba1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) or Novus (Littleton, CO, USA). The antibody for phospho-p47^phox (Ser370) was purchased from Assay Biotechnology Company Inc. (Sunnyvale, CA, USA). All other chemicals were obtained from Sigma–Aldrich, unless otherwise stated.

Lee E.J., Ko H.M., Jeong Y.H., Park E.M, & Kim H.S. (2015). β-Lapachone suppresses neuroinflammation by modulating the expression of cytokines and matrix metalloproteinases in activated microglia. Journal of Neuroinflammation, 12, 133.

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Effects of ASPS and Immune Challenge on Pig Growth

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Experiment (from June 5, 2012, to June 25, 2012) was conducted on the scientific pig farm of Shenyang Agriculture University (China). The formal feeding period lasted for 3 wks. The experiment was a 2×2 factorial arrangement including the main factors such as dietary ASPS supplementation (0 or 800 mg/kg) (Han et al., 2012a ) and immunological challenge (lipopolysaccharide [LPS] or saline injection). A total of 64 crossbred piglets (Duroc×Landrace×Large White), weaned at 28 d of age with average initial body weight (BW) of 7.25±0.21 kg were randomly assigned to 2 dietary groups with 8 replicate pens of 4 pigs per pen. On d 14, pigs in random four pens of per dietary treatment were intraperitoneally injected with 100 μg/kg BW LPS (Escherichia coli serotype 055:B5, Sigma) and the others were treated with equivalent amount of saline.
All pigs were housed in an environmentally controlled confinement house with concrete-slotted floor in 16 stainless steel pens (2.0×2.1 m). Each pen was equipped with a self-feeder and a nipple drinker to allow the pigs ad-libitum access to feed and water. The temperature in the inner house ranged from 21.6°C±2.3°C (6:00 to 7:00 am) to 23.2°C±1.5°C (15:00 to 16:00 pm) with 25.9°C±2.1°C on average. The basal diet containing no antibiotic, presented in Table 1, was formulated according to the nutrient requirements of NRC (1998) .

Han J., Bian L., Liu X., Zhang F., Zhang Y, & Yu N. (2014). Effects of Acanthopanax senticosus Polysaccharide Supplementation on Growth Performance, Immunity, Blood Parameters and Expression of Pro-inflammatory Cytokines Genes in Challenged Weaned Piglets. Asian-Australasian Journal of Animal Sciences, 27(7), 1035-1043.

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Inhibition of MMP and TACE Proteins

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LPS (Escherichia coli serotype 055:B5) was obtained from Sigma-Aldrich (St Louis, MO, USA). MMP-3 inhibitor (NNGH), MMP-8 inhibitor (M8I), and MMP-9 inhibitor (M9I) were purchased from Calbiochem (La Jolla, CA, USA). The chemical structures of the MMP inhibitors are illustrated in Fig. 1. The recombinant TACE protein was supplied by R&D Systems (Minneapolis, MN, USA). TAPI-0, recombinant MMP-3, MMP-8, and MMP-9 proteins were purchased from Enzo Life Sciences (Lausen, Switzerland). All cell-culture reagents were purchased from Gibco BRL (Grand Island, NY, USA). All other chemicals were obtained from Sigma-Aldrich, unless otherwise stated.

Lee E.J., Moon P.G., Baek M.C, & Kim H.S. (2014). Comparison of the Effects of Matrix Metalloproteinase Inhibitors on TNF-α Release from Activated Microglia and TNF-α Converting Enzyme Activity. Biomolecules & Therapeutics, 22(5), 414-419.

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Piglet Immune Response to LPS

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On the 14th day after birth, twelve piglets (three males and three females from each group) were selected in the study. One piglet per litter was selected, and the bodyweight of the selected piglets is close to the average bodyweight of the litter (Initial Weight: SD = 4.12 ± 0.05 kg vs FD = 4.11 ± 0.03 kg; p = 0.87). Blood samples (5 mL) were collected from each selected piglet, and then all selected piglets were administrated via the cervical side behind the left ear with E. coli LPS at 80 μg/kg BW. The LPS (Escherichia coli serotype 055: B5, Sigma Chemical, St. Louis, MO 63103, USA) was dissolved in sterile 0.9% NaCl solution (500 mg LPS per liter of saline). Blood samples (5 mL) were collected from each piglet at 5 h and 48 h post-LPS challenge. All piglets were weighed and recorded before slaughtering at 48 h post-LPS challenge. The internal organs (intestine, liver, kidney, spleen, heart and pancreas) of the piglets were obtained immediately after slaughtering and measured to calculate the relative organ weight and length to the bodyweight.

Luo W., Xu W., Zhang J., Yao J, & Xu J. (2020). The Maternal Diet with Fish Oil Might Decrease the Oxidative Stress and Inflammatory Response in Sows, but Increase the Susceptibility to Inflammatory Stimulation in their Offspring. Animals : an Open Access Journal from MDPI, 10(9), 1455.

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Acute Airway Inflammation Model in Mice

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A model of acute airway inflammation was elicited by the intranasal administration of lipopolysaccharide (LPS) (Escherichia coli serotype 055:B5, Sigma-Aldrich, St. Louis, MO, USA) inducing an inflammatory response with the extravasation of primarily PMNs in the airways (33 (link), 38 (link)). Specifically, male C57BL/6J wild-type (WT) and MPO-deficient MPO_tm1lus (MPO^-/-) mice (The Jackson Laboratory, USA), both aged 12-16 weeks and weighing 25-30 g, were subjected to brief anesthesia with ketamine-xylazine, after which 50 µl of LPS solution in phosphate-buffered saline (PBS) was instilled directly into the nostrils to achieve a dose of 0.3 mg/kg LPS, as described previously (33 (link), 38 (link)). Previous studies demonstrated that a significant fraction of intranasally administered LPS reaches the lungs, and that such instillation evokes an acute transient inflammatory response (33 (link), 38 (link)). All animal experiments were conducted in accordance with the EU Guide for the Care and Use of Laboratory Animals, and the experimental protocol was approved by the institutional Animal Care and Use Committee (The Czech Academy of Sciences of the Czech Republic, protocol n. 42/2015 from 12^th June 2015).

Kremserová S., Kocurková A., Chorvátová M., Klinke A, & Kubala L. (2022). Myeloperoxidase Deficiency Alters the Process of the Regulated Cell Death of Polymorphonuclear Neutrophils. Frontiers in Immunology, 13, 707085.

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