RASF and OASF were cultured as previously described [5] (link), and were used at passages 4–6. For TLR stimulation, 5×104 RASF per well in 6-well plates were incubated with different TLR ligands for 4 h and 24 h, respectively, as follows: TLR2 ligand peptidoglycan (PGN, 10 µg/ml), TLR3 ligand polyinosinic:polycytidylic acid (ploy(I:C), 25 µg/ml), TLR4 ligand lipopolysaccharide (LPS, 100 ng/mL) from Sigma; TLR9 ligand Type B CpG (10 µg/ml) from Invitrogen (Carlsbad, CA, USA). The cells were harvested for RT-PCR and realtime PCR while the supernatants were collected for ELISA.
Tlr4 ligand lipopolysaccharide lps
Manufactured by Merck Group
Sourced in United States
TLR4 ligand lipopolysaccharide (LPS) is a laboratory reagent used in research applications. It is a component of the outer membrane of Gram-negative bacteria and acts as a ligand for the Toll-like receptor 4 (TLR4). LPS can stimulate the innate immune response in cells expressing TLR4.
Automatically generated - may contain errors
Lab products found in correlation
Tlr9 ligand type b cpg, by Thermo Fisher Scientific (1 mentions)
Sb216763, by Bio-Techne (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
P38 inhibitor sb202190, by Bio-Techne (1 mentions)
Recombinant human il 1α and il 1β, by Thermo Fisher Scientific (1 mentions)
Nf κb inhibitor bay 11 7082, by Santa Cruz Biotechnology (1 mentions)
Jnk inhibitor sp600125, by Bio-Techne (1 mentions)
U0126, by Cell Signaling Technology (1 mentions)
3 protocols using tlr4 ligand lipopolysaccharide lps
1
TLR Stimulation of Fibroblast-Like Synoviocytes
2
Toll-Like Receptor Signaling Pathway
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TLR1/2 ligand Pam3CSK4, TLR2/6 ligand Peptidoglycan (PGN), TLR3 ligand polyinosinic-polycytidylic acid (Poly(I:C)), TLR5 ligand flagellin, were purchased from Invivogen (San Diego, CA, USA). Rabbit anti-human A20, ERK, β-actin, IκB-α, rabbit anti-human phosphorylated p38, ERK, JNK, were purchased from Cell Signaling Technology (Beverly, MA). TLR4 ligand lipopolysaccharide (LPS) was purchase from Sigma-Aldrich (St. Louis, MO). Scramble siRNA, human A20 siRNA, were purchased from Santa Cruz (Santa Cruz, CA). Human IL-1β, TNF-α, IL-8 ELISA kits were purchased from Jiamay Biotech. (Beijing, China). GSK3 selective inhibitor SB216763 was purchase from Tocris (Bristol, UK). A20 expressing plasmid was purchased from GeneCopoeia (Germantown, MD).
3
Signaling Pathway Modulation in Inflammation
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Rabbit anti-human A20 and GAPDH antibodies, rabbit anti-human phosphorylated ERK, JNK, p38, and NF-κB P65 antibodies, and the ERK inhibitor U0126, were purchased from Cell Signaling Technology (Beverly, MA, USA). The NOD2 ligand MDP was purchased from Invivogen (San Diego, CA, USA). The TLR4 ligand lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Human IL-8 and IL-1β ELISA kits were purchased from Jiamay Biotech. (Beijing, China). The A20 expression plasmid was purchased from GeneCopoeia (Germantown, MD, USA). The JNK inhibitor SP600125 and P38 inhibitor SB202190 were purchased from Tocris (Bristol, UK). The mouse anti-human NOD2 antibody and NF-κB inhibitor Bay 11-7082 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Recombinant human IL-1α and IL-1β were purchased from PeproTech Inc. (Rocky Hill, NJ, USA). NF-κB inhibitor BAY 11-7085 was purchased from MedChemExpress (Monmouth Junction, NJ, USA).
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