17β-Estradiol (E2), salicylamide (2-hydroxybenzamide), resorufin (7-hydroxy-3H-phenoxazin-3-one) and resorufin ethyl ether (7-ethoxy-3H-phenoxazin-3-one) were purchased from Sigma-Aldrich (Milan, Italy). G-1 (1-[4-(-6-bromobenzol [1 (link),3 (link)]diodo-5-yl)-3a,4,5,9b-tetrahidro3H5cyclopenta[c]quinolin-8yl]-ethanone), G-15 (3aS,4R,9bR)-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta[c]quinolone and TMS 1-[2,(3,5-Dimethoxyphenyl)ethenyl]-2,4-dimethoxybenzene were obtained from Tocris Bioscience (Space, Milan, Italy). Tyrphostin AG1478 (AG) and PD98059 (PD) were obtained from Calbiochem (DBA, Milan, Italy). All the aforementioned compounds were dissolved in dimethyl sulfoxide (DMSO), except for salicylamide that was dissolved in methanol.
Resorufin ethyl ether
Manufactured by Merck Group
Sourced in Italy, United States
Resorufin ethyl ether is a fluorescent dye used as a labeling agent in various biochemical and cell-based assays. It has an absorption maximum at 572 nm and an emission maximum at 585 nm, making it suitable for detection and quantification in fluorescence-based experiments.
Automatically generated - may contain errors
Lab products found in correlation
Resorufin, by Merck Group (3 mentions)
G 15 3as 4r 9br 4 6 bromo 1 3 benzodioxol 5 yl 3a 4 5 9b 3h cyclopenta c quinolone, by Bio-Techne (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
H4iie, by American Type Culture Collection (1 mentions)
Hepa1c1c7, by American Type Culture Collection (1 mentions)
2 3 7 8 tetrachlorodibenzo p dioxin, by Cambridge Isotopes (1 mentions)
Bradford reagent, by Merck Group (1 mentions)
Coumarin, by Merck Group (1 mentions)
Aminoguanidine, by Merck Group (1 mentions)
Umbelliferone, by Merck Group (1 mentions)
Nitrate reductase, by Merck Group (1 mentions)
Pentoxifylline ptx, by Merck Group (1 mentions)
4 protocols using resorufin ethyl ether
1
Assay of Estrogenic Compounds
2
TCDD and GNF-351 Cytotoxicity Assay
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2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, 50 μg/mL in DMSO, >99.9%) was purchased from Cambridge Isotope Laboratories (Tewksbury, MA) and GNF-351 was purchased from Santa Cruz Biotechnology (Dallas, TX). Trichloroacetic acid was purchased from Merck (Darmstadt, Germany). Resorufin ethyl ether, 3,3′-methylene-bis(4-hydroxycoumarin), sulforhodamine B and DMEM/F12 powder were purchased from Sigma (St Louis, MO). 100x L -glutamine and penicillin-streptomycin were purchased form Gibco (Dublin, Ireland) while fetal bovine serum was purchased from Cytiva Life Sciences (Marlborough, MA). HepG2, Hepa1c1c7 and H4IIE cell lines were purchased from ATCC (Manassas, VA).
3
Quantifying Nitric Oxide Production
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The following chemicals used in the experiments were obtained from Sigma Chemical Co. (St. Louis, MO, USA): pentoxifylline (PTX, P1784), aminoguanidine (AG, A7009), protein standard (P5619), Bradford reagent (B6916), resorufin (R3257), resorufin benzyl ether (B1532), resorufin ethyl ether (E3763), umbelliferone (U7626), coumarin (C4261), nitrate reductase (N7265), reduced nicotinamide adenine dinucleotide phosphate (β-NADPH, N1630), sodium nitrate (S5506), N-(1-naphthyl)ethylenediamine (N9125), sulphanilamide (S9251), nicotinamide adenine dinucleotide phosphate (β-NADP, N0505), glucose-6-phosphate (G7250), glucose-6-phosphate dehydrogenase (G6378) and Escherichia coli LPS (type 0127:B8, L3129). All other chemicals used were of high analytical grade.
4
CYP1A1 Activity Assay in BMDE Cells
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BMDEo from day 14 of culture were seeded at 1 × 106 cells per 300 μl in 96-well plates and cultured with the indicated concentration of FICZ for 4 h. Cells were washed in PBS and resuspended in 100 μl PB (50 mM NaHPO4 and 50 mM NaH2PO4, pH 8.0) with 2 µM resorufin ethyl ether (#E3763; Sigma-Aldrich) and incubated for 30 min. resorufin ethyl ether is converted to resorufin by the CYP1A1 enzyme. The reaction was stopped by addition of 75 μl acetonitrile containing fluorescamine (150 µg/ml). resorufin fluorescence was measured at 535-nm excitation and 590-nm emission. Fluorescamine fluorescence was measured at 390-nm excitation and 485-nm emission. Serial dilutions of resorufin (#424455; Sigma-Aldrich) and BSA were measured in parallel to generate standard curves. resorufin concentration was normalized to protein content.
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