1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), 1,2-dipalmitoleoyl-sn-glycero-3-phosphoethanolamine (DiPoPE), 1-palmitoyl-2-linoleoyl-sn-phosphatidylcholine (PLPC) and stearylamine were purchased from Avanti Polar Lipids, Inc. (Alabaster, Alabama). Ammonium formate, propanal, butanal, pentanal, hexanal, octanal, nonanal and 7-(diethylamino)-coumarin-3-carbohydrazide (CHH) were supplied by Sigma-Aldrich GmbH, and sodium cyanoborohydride (NaCNBH4) was purchased from FlukaChemie GmbH (Buchs, CH). Ultra liquid chromatography grade acetonitrile and methanol were from Biosolve BV (Valkenswaard, Netherlands). Ethanol and acetic acid were from Carl Roth GmbH & Co. KG and chloroform was from Merck KGaA (Darmstadt, Germany). Peptide GGQEHFAHKLILR (N-terminus acetylated) was synthesized in-house by solid phase peptide synthesis on a multiple peptide synthesizer (SYRO 2000, MultiSynTech GmbH, Witten, Germany) using Fmoc-chemistry. l -glutamine, penicillin, streptomycin, lipopolysaccharide, peptidoglycan from Staphylococcus aureus, phorbol 12-myristate 13-acetate and ionomycin were purchased from Sigma-Aldrich GmbH.
Peptidoglycan from staphylococcus aureus
Manufactured by Merck Group
Sourced in United States
Peptidoglycan from Staphylococcus aureus is a component of the bacterial cell wall. It is a complex molecule composed of amino acids and sugars that provides structural support and protection for the bacterial cell.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Ammonium formate, by Merck Group (1 mentions)
Octanal, by Merck Group (1 mentions)
Nonanal, by Merck Group (1 mentions)
Hexanal, by Merck Group (1 mentions)
Lipopolysaccharide, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
7 diethylamino coumarin 3 carbohydrazide, by Merck Group (1 mentions)
Propanal, by Merck Group (1 mentions)
Pentanal, by Merck Group (1 mentions)
1 2 dipalmitoyl sn glycero 3 phosphoethanolamine, by Avanti Polar Lipids (1 mentions)
Butanal, by Merck Group (1 mentions)
1 2 dipalmitoleoyl sn glycero 3 phosphoethanolamine, by Avanti Polar Lipids (1 mentions)
Methanol, by Biosolve (1 mentions)
Acetic acid, by Carl Roth (1 mentions)
Escherichia coli 0111 b4, by Merck Group (1 mentions)
4 protocols using peptidoglycan from staphylococcus aureus
1
Lipid Profiling and Peptide Synthesis Protocol
2
TLR-Agonist Stimulation of Immune Cells
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SCC24, SCC90, SCC72, SCC89 were stimulated with various concentrations of lipopolysaccharide (LPS: a non-specific TLR4 agonist) from Escherichia coli 0111:B4 (Sigma-Aldrich, St. Louis, MO, USA), lipopolysaccharide ultra-pure (LPS-UP: a specific TLR4 agonist) from E. coli (In vivo Gen, San Diego, California, USA), and peptidoglycan from Staphylococcus aureus (PGN: a TLR2 agonist) (Sigma-Aldrich, St. Louis, MO, USA). For ELISA, cells were stimulated with 0.01–1 µg/mL of LPS; 0.01-1 µg/mL of LPS-UP and 0.1–10 µg/mL of PGN [25 (link), 56 (link)]. For qPCR and Western blot, cells were stimulated with 1 µg/mL of LPS or LPS-UP or PGN. All incubations were undertaken in a humidified incubator (5% CO2) at 37°C for 24 h. Experiments were undertaken using triplicate biological repeats.
3
Quantifying Inflammatory Mediators in Cells
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Peptidoglycan (from Staphylococcus aureus, lot number 77140), lipopolysaccharide (LPS), thrombin, and bradykinin (BK) were purchased from Sigma-Aldrich Chemical Co. (St Louis, MO, USA). The COX-1 antibody was obtained from Santa Cruz Biotechnology Inc. (Dallas, TX, USA), whereas the COX-2 antibody was purchased from BD Biosciences (Franklin Lakes, NJ). The β-actin antibody was purchased from EMD Millipore (Billerica, MA, USA). U46619 (a TXA2 analog), PGE2, AH6809, and AH23848 were purchased from Cayman Chemicals (Ann Arbor, Michigan, USA). α-SMA antibody was purchased from GeneTex Inc. (Hsinchu, Taiwan).
4
Immune Cell Culture and TLR Ligand Preparation
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RAW G9 cells were maintained in DMEM, 10% FBS, 20 mM Hepes, and 2 mM glutamine. THP1 B5 cells were maintained in RPMI1640, 10% FBS, and 2 mM glutamine containing 500 ug/ml G418. TLR ligand sources: LPS was from Alexis Biochemicals, Salmonella minnesota R595 TLRgrade, ALX-581-008-L002; Pam3CSK4 (P3C) was from EMC Microcollections, cat# L2000; PGN was from Sigma, peptidoglycan from Staphylococcus aureus, Cat.# 77140; R848 was from InvivoGen, tlrl-r848; Pam2CSK4 (P2C) was from EMC Microcollections, cat# L2020; Flagellin was from Invivogen, FLA-ST ultrapure, tlrl-epstfla; Lipid A was from Avanti Polar Lipids, 699500P; CpG was from IDT; poly I:C was from Invivogen, tlrl-picw.
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