HPLC-grade acetonitrile was purchased from Tedia (Fairfield, OH, USA). Amino acid standards, ABTS, DPPH, 2,4,6-tripyridyl-s-triazine (TPTZ), LPS, CCK-8, rat IL-1β, PGE2, and the TNF-α ELISA kit were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), and 1% penicillin/streptomycin were bought from Gibco (Carlsbad, CA, USA). The synthesized peptides (purity ≥ 95%) were obtained from Bankpeptide Biological Technology Co., Ltd. (Hefei, China). Other reagents were of analytical grade.
Rat il 1β
Manufactured by Merck Group
Sourced in United States
Rat IL-1β is a laboratory product that serves as a recombinant cytokine. It is a member of the interleukin-1 family and plays a role in the regulation of immune and inflammatory responses.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Cck 8, by Merck Group (1 mentions)
Tnf α elisa kit, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
2 4 6 tripyridyl s triazine, by Merck Group (1 mentions)
Hplc grade acetonitrile, by Tedia (1 mentions)
Celltiter96 assay, by Promega (1 mentions)
Rat ifn γ, by Thermo Fisher Scientific (1 mentions)
Staining kit, by Beyotime (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
P iκbα, by ABclonal (1 mentions)
Tgf β1, by ABclonal (1 mentions)
Adamts 4, by ABclonal (1 mentions)
Ex 527, by Merck Group (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Smad2, by ABclonal (1 mentions)
Cox 2, by ABclonal (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
4 protocols using rat il 1β
1
Antioxidant and Anti-inflammatory Assays
2
Thapsigargin and Cytokine-Induced Cell Toxicity
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832/13 cells were seeded in 96-well tissue culture dishes. To induce cell toxicity, cells were treated with 200–500 nM thapsigargin or with a mixture of 1 ng/ml rat IL-1β (Sigma) + 100 U/ml rat IFN-γ (Thermo). Cell viability was measured using the CellTiter96 assay (Promega), and activation of apoptosis was measured by caspase-3 cleavage [16 ].
3
Inflammatory Cytokine Regulation in Rat Cells
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MT, Rat IL-1β, 4′,6-diamidino-2-phenylindole (DAPI), collagenase type II, and EX527 were purchased from Sigma Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS) (BI, Kibbutz Bet Hamek, Israel), Dulbecco’s Modified Eagle’s Medium-(DMEM-) F12 Medium, and Phosphate Buffered Saline purchased from Corning (New York, NY, USA). 1% penicillin/streptomycin, cell counting kit-8 (CCK-8), and staining kit were purchased from Beyotime (Haimen, China). Trypsin (0.25%) was purchased from Gibco (Grand Island, NY, USA). Lysate and RAPI were purchased from Biyuntian (Shanghai, China). nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p65), Phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65),alpha (IκBα), p-IκBα, TGF-β1, Smad2, iNOS, COX-2, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), collagen type II alpha 1 chain (COL2A1), SIRT1, and ADAMTS-4 were purchased from ABclonal Technology (Wuhan, China). Horseradish peroxidase-conjugated goat anti-mouse/rabbit immunoglobulin G (IgG) secondary antibody was purchased from ABclonal Technology (UK). GAPDH was purchased from CST (Danvers, MA, USA).
4
Investigating Blood-Brain Barrier Inflammation
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The RBMECs were purchased from Procell (Wuhan, China). These cells, which were isolated from brain tissues, are the main component of the blood-brain barrier. After the RBMECs were thawed in warm water at 37°C, they were cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM; Gibco, Waltham, USA) containing 10% fetal bovine serum (FBS), 100 U/mL of penicillin and 100 mg/mL of streptomycin (Gibco) at 37°C in 5% CO 2 . The medium was replaced every 2 days and the cells selected for the experiments were all in the 3 rd to 6 th generation. To simulate an inflammatory environment, RBMECs were treated with rat IL-1β (1 ng/mL; Sigma-Aldrich, St. Louis, USA) for 3 h. After IL-1β treatment, the RBMECs were treated with the ERK1/2 specific upstream inhibitor PD98059 (50 μM; MedChemExpress, Monmouth Junction, USA) for 1 h. PD98059 is an effective inhibitor of the MEK signaling pathway through its blockade of MEK1 and MEK2 with an IC 50 of 5 µM. It also inhibits the ERK1/2 signaling pathway by blocking phosphorylation. In preparation for the experiments, the cells were grouped and named the negative control (NC) group, the IL-1β group and the PD98059 group.
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