Lipopolysaccharide

Manufactured by Solarbio

Sourced in China

Lipopolysaccharide is a complex molecule found in the outer membrane of Gram-negative bacteria. It serves as a major structural component of the bacterial cell wall. Lipopolysaccharide is commonly used in research and laboratory settings to study the immune response and bacterial pathogenesis.

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11 protocols using lipopolysaccharide

Mouse Macrophage Inflammatory Response

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Mouse macrophages (RAW264.7 cells) were purchased from Shanghai Cell Bank, Chinese Academy of Sciences. AEE (99.5%) was obtained from Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS. (Lanzhou, China). Asp and Eug were supplied by Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Lipopolysaccharide, trypsin, and Cell Counting Kit-8 were from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Foundation FBS and DMEM high glucose medium were purchased from MCE Co., Ltd. (Shanghai, China). ELISA kits including TNF-α (JL10484), IL-1β (JL18442), IL-6 (JL20268) and IL-8 (JL20271) were supplied by Shanghai Jiang Lai Biotechnology Co., Ltd. (Shanghai, China). RNA extraction kit, reverse transcription kit, fluorescence quantitative PCR kit were from Mei5 Biotech Co., Ltd. (Beijing, China). β-actin (3700s), p65 (8242s), p-p65 (3033s), p38 (8690s), p-p38 (4511s) were obtained from Cell Signaling Technology., Inc. (Danvers, MA, United States). The isotope internal standards including TXB₂-d4 (319030), PGE₂-d9 (10581), PGF_2α-d4 (316010), 6-keto-PGF_1α-d4 (15210), PGD₂-d4 (312010), 9-HODE-d4 (338410), 13-HODE-d4 (338610), 12-HETE-d8 (334570), 15-HETE-d8 (334720), DHA-d5 (10005057), AA-d8 (390010) were purchased from Cayman Chemical Co., (Ann Arbor, MI, United States).

Liu X., Tao Q., Shen Y., Liu X., Yang Y., Ma N, & Li J. (2023). Aspirin eugenol ester ameliorates LPS-induced inflammatory responses in RAW264.7 cells and mice. Frontiers in Pharmacology, 14, 1220780.

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Evaluating Anti-Inflammatory Activity of Films

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Anti-inflammatory response of films was assessed using RAW 264.7 cells (leukemia cells in mouse macrophage cell line). Briefly, cells (1 × 10⁶ cells/mL) were seeded onto sterilized films in 24-well plates in a humidified incubator (37 °C, 5% CO₂) using complete Dulbecco's Modified Eagle's medium (DMEM, Gibco, USA) media. In order to assess the anti-inflammatory activity, RAW 264.7 cells were stimulated with 10 ng/mL LPS (Lipopolysaccharide, Solarbio, China) compared with the blank control (tissue culture plastic). After 18 h, cells were collected to analyze TNF-α and IL-1β expression by RT-qPCR. The morphology of cells was evaluated using SEM after rinsed with PBS, dehydrated through graded ethanol, and dried in a vacuum. Viability of cells on films were also observed after labeled with Live/Dead reagent.

Zheng M., Guo J., Li Q., Yang J., Han Y., Yang H., Yu M., Zhong L., Lu D., Li L, & Sun L. (2020). Syntheses and characterization of anti-thrombotic and anti-oxidative Gastrodin-modified polyurethane for vascular tissue engineering. Bioactive Materials, 6(2), 404-419.

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Evaluating the Immunomodulatory Potential of Abrus cantoniensis

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Abrus cantoniensis were provided by the Guangxi Dahong Pharmaceutical Co., Ltd. (Hechi, Guangxi, China). After being crushed and sun-dried, the material was stored at 4°C.
Dimethyl sulfoxide (DMSO), DEAE cellulose-52, lipopolysaccharide (LPS), ROS assay Kit, RPMI 1640 medium and neutral red were acquired from Solarbio Biotechnology Co., Ltd. (Beijing, China). Monosaccharide standards (glucose, mannose, arabinose, galactose, fucose, rhamnose, glucuronic acid, galacturonic acid, glucosamine, Galactosamine, xylose), trifluoroacetic acid (TFA), 1-phenyl-3-methyl-5-pyrazolone (PMP) were purchased from Sigma (St. Louis, MO, United States). The Vybrant phagocytosis assay kit (V-6694) was obtained from Thermo Fisher Scientific (Waltham, United States). The ELISA kits for mouse tumor necrosis factor (TNF-α), interferon (IFN-γ), interleukin (IL-6, IL-1β) were provided by Jiangsu Jingmei Biotechnology Co., Ltd (Jiangsu, China). The NO and Inducible nitric oxide synthase (iNOS) detecting kit were acquired from Nanjing Jiancheng Institute of Biotechnology (Nanjing, China). Trizol reagent, RealStar Green Fast Mixture and First-strand cDNA Synthesis Mix were bought from GenStar Co., Ltd. (Beijing, China).

Qu D., Lian S., Hu H., Sun W, & Si H. (2022). Characterization and macrophages immunomodulatory activity of two water-soluble polysaccharides from Abrus cantoniensis. Frontiers in Nutrition, 9, 969512.

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Murine Pancreatitis Model Exploration

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Male C57BL/6 J mice from Beijing Vital River (China) were used in this study. The AR42J cell line and F-12 K medium were obtained from the BeNa culture collection (China). Recombinant mouse Interleukin -22 (Miltenyi, Germany) was used, along with Caerulein (Sigma, America) and Lipopolysaccharide (Solarbio, China). AKT, mTOR, anti-mTOR (phosphor s2448), and anti-AKT (Ser473) antibodies were purchased from Proteintech(China). LC3 and P62 antibodies were obtained from Abclonal(China), and GAPDH Rabbit Polyclonal antibody and goat anti-rabbit IgG(H + L) HRP conjugate were from Proteintech(China). Trizol reagent, PCR primers, Reverse transcription kit, and SYBR Green qPCR kit were purchased from Agbio(China).

Fu X., Xiu Z., Xu Q., Yue R, & Xu H. (2024). Interleukin-22 Alleviates Caerulein-Induced Acute Pancreatitis by Activating AKT/mTOR Pathway. Digestive Diseases and Sciences, 69(5), 1691-1700.

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Bone Marrow-Derived Dendritic Cell Protocol

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LBP was purchased Shanxi Ciyuan Biotechnology (No.CY190218). Fetal bovine serum was purchased from PNA, Paisley, UK. Recombinant Murine GM-CSF and recombinant Murine IL-4 were both from PeproTech (USA). Lipopolysaccharide was from Solarbio, China.

Duan X., Lan Y., Zhang X., Hou S., Chen J., Ma B., Xia Y, & Su C. (2020). Lycium barbarum Polysaccharides Promote Maturity of Murine Dendritic Cells through Toll-Like Receptor 4-Erk1/2-Blimp1 Signaling Pathway. Journal of Immunology Research, 2020, 1751793.

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Investigating SR9009 Protective Effects on Inflammatory Injury

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The NPMSC were incubated with a graded concentration of SR9009 (Solarbio, Beijing, catalog no. SG8180) (0–40 μM) to evaluate the protective effect of SR9009 on Lipopolysaccharide/Adenosine Triphosphate-induced inflammatory injury. Based on the data showed in “Results” section, NPMSC were prior to be incubated with 10 μM SR9009 for 12 h and then 1 μg/ml Lipopolysaccharide (Nan Jing, Jian cheng, catalog no. E004) for 24 h and 5 mM ATP for last half hour as the intervention for the follow-up experiment according to Wang et al. description for inflammation model.²⁷ (link) The cells were divided into four groups according to different treatment: (A) CON group: blank control; (B) LPS+ATP group: 1 μg/ml Lipopolysaccharide+5mM Adenosine Triphosphate; (C) SR9009 group: 10 μM SR9009; (D) S + L + A group: 10 μM SR9009 + 1 μg/ml Lipopolysaccharide + 5mM Adenosine Triphosphate; (E) YVAD group: 10 μM YVAD + 1 μg/ml Lipopolysaccharide +5 mM Adenosine Triphosphate.

Huang Z.N., Wang J., Wang Z.Y., Min L.Y., Ni H.L., Han Y.L., Tian Y.Y., Cui Y.Z., Han J.X, & Cheng X.F. (2024). SR9009 attenuates inflammation-related NPMSC pyroptosis and IVDD through NR1D1/NLRP3/IL-1β pathway. iScience, 27(5), 109733.

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Phenolic Compounds for BV2 Cell Activation

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Ethylene glycol dimethyl methacrylate, 4-ethylene pyridine, and silane coupling agent KH-570 were purchased from Aladdin Reagent Co., Ltd. (Shanghai, China). Azo diisobutyronitrile was acquired from the Tianjin Guangfu Fine Chemical Research Institute (Tianjin, China). Acrylamide (AM) was purchased from Shanpu Chemical Co., Ltd. (Shanghai, China). The silica gel (30 to 40 μm) was supplied by Dingkang Silica Gel Co., Ltd. (Qingdao, China). Six phenolic compounds (ellagic acid, kaempferol-3-o-rutinoside, gallic acid, vanillic acid, ferulic acid and tiliroside) were purchased from ERFA Biotechnology Co., Ltd. (Chengdu, China). BV2 cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China). Lipopolysaccharide, 0.25% trypsin-EDTA solution, and high glucose medium DMEM were obtained from Solarbio Co., Ltd. (Beijing, China). Fetal bovine serum was procured from Procell Company (America). PBS was supplied by Biochem Co., Ltd. (Shenzhen, China). The CCK8 reagent test kit was purchased from KGI Bio (Suzhou, China). The ELISA kit was purchased from Enzyme Immunoassay Industrial Co., Ltd. (Suzhou, China).

Wu Q., Naeem A., Zou J., Yu C., Wang Y., Chen J, & Ping Y. (2022). Isolation of Phenolic Compounds from Raspberry Based on Molecular Imprinting Techniques and Investigation of Their Anti-Alzheimer’s Disease Properties. Molecules, 27(20), 6893.

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Curcumin and Lipopolysaccharide Protocol

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Curcumin (purity > 98%, 08511-10MG) was purchased from Sigma (Chemical Co., St. Louis, MO, USA), and lipopolysaccharide (purity ≥ 99%, L8880-10 mg) was purchased from Solarbio (Beijing, China).

Liu Y., Wang Z., Gan Y., Chen X., Zhang B., Chen Z., Liu P., Li B., Ru F, & He Y. (2021). Curcumin attenuates prostatic hyperplasia caused by inflammation via up-regulation of bone morphogenetic protein and activin membrane-bound inhibitor. Pharmaceutical Biology, 59(1), 1024-1033.

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Capsaicin and Lipopolysaccharide in Mice

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The mice were randomly allocated into four groups (n=6 per group): (1) Control group (CON); (2) lipopolysaccharide-treated group (LPS); (3) capsaicin diet group (CAP); (4) capsaicin diet + lipopolysaccharide-treated group (CAP+LPS). The mice of Control and LPS groups were fed standard laboratory chow (Department of Laboratory Animal Science of China Medical University). The mice in the CAP and CAP+LPS groups were fed standard laboratory chow plus 0.005% capsaicin (Targetmol, Boston, MA, USA). All mice were fed for 4 months. During the last 5 days, mice in LPS and CAP+LPS groups received lipopolysaccharide (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) once daily via intraperitoneal injection. The following doses of LPS were used according to previous methods: 0.052/0.104/0.208/0.415/0.83 mg/kg (5-day) (Wickens et al., 2017 (link)). The CON and CAP groups were injected with an equal volume of saline in the same manner. Body weight was measured after behavioral testing.

Xia J., Gu L., Guo Y., Feng H., Chen S., Jurat J., Fu W, & Zhang D. (2021). Gut Microbiota Mediates the Preventive Effects of Dietary Capsaicin Against Depression-Like Behavior Induced by Lipopolysaccharide in Mice. Frontiers in Cellular and Infection Microbiology, 11, 627608.

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Endothelial Cell Isolation and Characterization

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EBM-2 culture medium (Lonza, USA), ficoll hypaque 1.077 (Haoyang, Tianjin, China), lipopolysaccharide (Solarbio, Beijing, China), CCK-8 kit (Dojindo Laboratories, Japan), FITC-coupled annexin-V apoptosis detection kit (BD Biosciences, USA), cell cycle and apoptosis kit (Beyotime Biotechnology, Shanghai, China), FITC-UEA-I/ DiI-acLDL (Proteintech, USA), PCDH-CMV-EF1-copGFP-Sirt1 (LabGene, Guangzhou, China), SRT1720/EX527 (Sigma-Aldrich, USA), reverse transcription kit (Thermo Fisher, USA), rabbit monoclonal antibodies to Sirt1/β-actin (ab189494/ ab115777, Abcam, UK), rabbit monoclonal antibodies to p53/NF-κB/FOXO3a (32532S/8242S/12829S, Cell Signaling, USA), HRP conjugated AffiniPure goat anti-rabbit (BA1055, BOSTER, Wuhan, China), PCR fluorescence quantification kit (Thermo Fisher, USA), Matrigel™ Matrix (BD Biosciences, USA), and CD34+ positive selection cocktail (EasySep, Stemcell, Canada) were used in the experiments.

Sun Y., Liu X., Sun D., Yao J., Zhan-ling D., Qian J., & Huang Q. (2021). Effects of Sirt1 on proliferation, migration, and apoptosis of endothelial progenitor cells in peripheral blood of SD rats with chronic obstructive pulmonary disease. Asian Pacific Journal of Tropical Biomedicine, 11(10), 429-429.

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