Endotoxin shock was induced by intravenous injection of LPS (Escherichia coli O111:B4; L2630, Sigma-Aldrich, St. Louis, MO, USA) dissolved in 0.2 mL of normal saline at a dose of 10 mg/kg 28 (link), 29 . The animals were continuously observed for 48 hours after LPS administration and sacrificed thereafter.
Lps escherichia coli o111 b4 l2630
Manufactured by Merck Group
Sourced in United States, China
LPS (Escherichia coli O111:B4; L2630) is a laboratory product supplied by Merck Group. It is a lipopolysaccharide (LPS) derived from the Escherichia coli O111:B4 strain. LPS is a major component of the outer membrane of Gram-negative bacteria and is commonly used in research applications.
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Lab products found in correlation
Anti gsdmd ab209845, by Abcam (1 mentions)
Anti caspase 1 ag 20b 0042, by Adipogen (1 mentions)
Poly da dt tlrl patn, by InvivoGen (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Anti brd4 a301 985a, by Fortis Life Sciences (1 mentions)
Il 6 88 7064 22 elisa kits, by Thermo Fisher Scientific (1 mentions)
Nigericin n7143, by Merck Group (1 mentions)
Anti asc ag 25b 0006 c100, by Adipogen (1 mentions)
Anti h3k27ac ab4729, by Abcam (1 mentions)
Acetonitrile, by CNW Technologies (1 mentions)
Formic acid, by CNW Technologies (1 mentions)
Methanol, by CNW Technologies (1 mentions)
Sb 505124, by Merck Group (1 mentions)
4 protocols using lps escherichia coli o111 b4 l2630
1
Endotoxin Shock Induction Protocol
2
LPS, Nigericin, and DOTAP Modulation of Inflammatory Responses
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LPS (Escherichia coli O111:B4, L2630), nigericin (N7143) and N-(2,3-dioleoyloxy-1-propyl)trimethylammonium methyl sulfate (DOTAP) liposomal transfection reagent (144189-73-1) were from Sigma-Aldrich. Ultrapure flagellin from S. typhimurium (tlrl-epstfla-5) and poly(dA:dT; tlrl-patn) were purchased from InvivoGen. TcdB from List Labs (155). Lipofectamine RNAiMAX Transfection Reagent (13778150) was from Invitrogen. Mouse TNF-α (88-7324-88), IL-1β (88-7013-88), IL-18 (BMS618-3), and IL-6 (88-7064-22) ELISA kits were from Invitrogen. The LightShift Chemiluminescent EMSA Kit (20148) was from Thermo Fisher Scientific Inc. Anti–IL-1β (AF-401-NA) was from R&D Systems. Anti–caspase-1 (AG-20B-0042) and anti-ASC (AG-25B-0006-C100) were from AdipoGen. Anti-GSDMD (ab209845) and anti-H3K27ac (ab4729) were from Abcam. Antiactin (sc-47778), anti-IRF8 (sc-365042), anti-RNAPII (sc-47701), and anti-PU.1 (sc-390405) were from Santa Cruz Biotechnology, Inc. Anti-Brd4 (A301-985A) was from Bethyl Laboratories. Anti-H3K4me1 (07-436) and anti-H3K4me3 (05-745R) were from EMD Millipore.
3
Analytical Grade Chemicals Protocol
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All chemicals and solvents were analytical or HPLC-grade. Acetonitrile, methanol, and formic acid were purchased from CNW Technologies GmbH (Düsseldorf, Germany). L-2-chlorophenylalanine was obtained from Shanghai Hengchuang Biotechnology Co., Ltd. (Shanghai, China), and LPS (Escherichia coli (O111:B4), #L2630, purity >99%, with high-purity quality Standards M300) was commercially available from Sigma-Aldrich (Saint Louis, MO, United States).
4
Labrum Explant Culture for Inflammatory Modeling
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Explant culture of human labrum tissues was adapted from a method developed previously for osteochondral joint tissues [19 (link)]. Clinical specimens were processed within 2 h post-surgery and dissected with a scalpel into equal-sized samples (200–400 mg wet weight). Dissected tissues were cultured in 8 mL αMEM (10% FBS, 10 mM HEPES, 4 mM l -glutamine) for 1 week at 37 °C in a humidified atmosphere containing 5% CO2. Specimens were treated with 1 μg/mL LPS (Escherichia coli O111:B4, L2630 Sigma–Aldrich) in the presence of a TGF-beta receptor type I inhibitor (10 μM SB-505124) or DMSO vehicle (0.00075% v/v) at day 1 and 4. Conditioned medium was collected at day 7 and stored at −80 °C until further analysis. Labrum tissues were fixed in formalin for 48 h at 4 °C.
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