Cells were treated for 48 h beginning at day 2 of differentiation with DMSO (vehicle control, ATCC), aceclidine (Sigma-Aldrich), acyclovir (Sigma-Aldrich), alloxazine (Sigma-Aldrich), carbadox (Sigma-Aldrich), felodipine (Sigma-Aldrich), GW5074 (Sigma-Aldrich), isoproterenol (Sigma-Aldrich), isradipine (Sigma-Aldrich), lacidipine (Sigma-Aldrich), nafadotride (Tocris Bioscience), nandrolone (Sigma-Aldrich), nifedipine (Sigma-Aldrich), nilvadipine (Sigma-Aldrich), alloxazine (Sigma-Aldrich) diluted in cell-type specific differentiation media at doses listed in figures.
Alloxazine
Manufactured by Merck Group
Sourced in United States, Italy
Alloxazine is a chemical compound used in various laboratory applications. It serves as a functional component in certain types of lab equipment and instrumentation. The core function of Alloxazine is to facilitate specific chemical reactions and processes within controlled laboratory environments.
Automatically generated - may contain errors
Lab products found in correlation
Carbadox, by Merck Group (1 mentions)
Acyclovir, by Merck Group (1 mentions)
Isradipine, by Merck Group (1 mentions)
Nifedipine, by Merck Group (1 mentions)
Felodipine, by Merck Group (1 mentions)
Lacidipine, by Merck Group (1 mentions)
Nandrolone, by Merck Group (1 mentions)
Isoproterenol, by Merck Group (1 mentions)
Gw5074, by Merck Group (1 mentions)
Phytohemagglutinin (pha), by Merck Group (1 mentions)
N nitro l arginine methyl ester l name, by Merck Group (1 mentions)
Hemin, by Merck Group (1 mentions)
3h thymidine, by GE Healthcare (1 mentions)
Indomethacin, by Merck Group (1 mentions)
Acetonitrile, by Avantor (1 mentions)
Alloxan, by Merck Group (1 mentions)
Ptfe filter, by Thermo Fisher Scientific (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
O phenylenediamine, by Merck Group (1 mentions)
4 protocols using alloxazine
1
Dose-Dependent Differentiation Modulation
2
Immunomodulatory Effects of MSCs
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Human PBMCs (1 × 105/well) from healthy adult donors were plated in 96-well flat-bottomed plates (200 μL/well). Cells were then stimulated with 5 μg/mL phytohemagglutinin (Sigma-Aldrich) and/or MSCs (5 × 103, 1 × 104, 5 × 104, 1 × 105) for 72 h, followed by the incorporation of 1 μCi/mL [3H]-thymidine (GE Healthcare, Piscataway, NJ, United States) for the last 18 h of the indicated culture period. Radioactivity was measured using a Micro Beta instrument (Pharmacia Biotech, Piscataway, NJ, United States).
In other experiments, neutralizing antibodies for human IL-10 or transforming growth factor-β mAb (10 μg/mL; R and D Systems) and chemical antagonists for COX2 (indomethacin, 20 μM; Sigma-Aldrich), iNOS [N-nitro-L-arginine methyl ester (L-NAME), 1 mM; Sigma-Aldrich], heme oxygenase 1 inducer (hemin, 50 ng/mL; Sigma-Aldrich), selective A2B adenosine receptor (alloxazine, 10 μM; Sigma-Aldrich), and CD73 (α,β-methylene ADP [APCP], 100 µM; Sigma-Aldrich) were added into the coculture. All experiments were performed in quadruple and were repeated at least twice.
In other experiments, neutralizing antibodies for human IL-10 or transforming growth factor-β mAb (10 μg/mL; R and D Systems) and chemical antagonists for COX2 (indomethacin, 20 μM; Sigma-Aldrich), iNOS [N-nitro-L-arginine methyl ester (L-NAME), 1 mM; Sigma-Aldrich], heme oxygenase 1 inducer (hemin, 50 ng/mL; Sigma-Aldrich), selective A2B adenosine receptor (alloxazine, 10 μM; Sigma-Aldrich), and CD73 (α,β-methylene ADP [APCP], 100 µM; Sigma-Aldrich) were added into the coculture. All experiments were performed in quadruple and were repeated at least twice.
3
Adenosine Receptor Inhibition Modulates Cytokine Expression
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The A2b receptor inhibitor alloxazine (Sigma–Aldrich) was added at a concentration of 100 nm to the medium for pharmacological inhibition of adenosine receptors. After 2 days, culture medium was replaced. Next, the expressions of IL‐10 and IL‐1RA genes were determined using qRT‐PCR following the method mentioned above. Moreover, immunofluorescence staining was used to determine the expression level of Arg‐1 protein.
4
Analytical Method for Alloxan and Alloxazine
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Acetonitrile and formic acid 99.9% (LC-MS grade), water (HPLC gradient grade), were supplied from VWR (VWR International PBI Srl Milan, Italy). Alloxan, Alloxazine, o-phenylenediamine, hydrochloride acid 1M and sodium hydroxide 1M were purchased from Sigma (Sigma-Aldrich, Milan, Italy). Standard stock solutions of Alloxan and o-phenylenediamine were prepared in hydrochloride acid 0.1M (concentration of 1 mg mL -1 ). Standard stock solution of Alloxazine was prepared in sodium hydroxide 0.1M at the concentration of 1 mg mL -1 .These solutions were stored at 2 to 8 ºC up to 3 months. Working stock solutions of Alloxan and Alloxazine were prepared in formic acid 0.1% (v/v) in water at a concentration of 10 µg mL -1 and stored at 2 to 8 ºC up to 30 days. PTFE filters of 0.45 µm were purchased from Chromacol LTD (Thermo Fisher, Waltham, Massachusetts, USA).
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