Analytical-grade reagents and reference substances were obtained from Aldrich (Milwaukee, MN, USA). Phenobarbital (PB) was purchased from Abbott Laboratories (Mexico City, Mexico). Beta-naphthoflavone (β-NF), resorufin, 7-ethoxyresorufin (ER), methoxyresorufin (MR), benzyloxiresorufin (BR), pentoxyresorufin (BR), NADPH, BP, 2,2-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), thiobarbituric acid (TBA), corn oil, and Giemsa stain were purchased from Sigma Chemicals Co. (St. Louis, MO, USA). Microsomes of Baculovirus expression systems from rat CYP1A1-expressing insect cells (Supersomes) were purchased from BD-Gentest (Woburn, MA, USA).
Methoxyresorufin
Manufactured by Merck Group
Sourced in United States
Methoxyresorufin is a fluorescent dye compound used in laboratory settings as a substrate for the detection and quantification of cytochrome P450 enzymes, particularly CYP1A. It is a selective and sensitive probe for these enzymes, which play a crucial role in the metabolism of various substances. Methoxyresorufin can be used in a variety of analytical techniques, such as fluorescence-based assays, to measure the activity of CYP1A enzymes in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ethoxyresorufin, by Merck Group (13 mentions)
Resorufin, by Merck Group (12 mentions)
Nadph, by Merck Group (9 mentions)
P nitrophenol, by Merck Group (7 mentions)
Erythromycin, by Merck Group (6 mentions)
Pentoxyresorufin, by Merck Group (5 mentions)
Benzyloxyresorufin, by Merck Group (5 mentions)
1 chloro 2 4 dinitrobenzene, by Merck Group (5 mentions)
7 ethoxyresorufin, by Merck Group (4 mentions)
Glucose 6 phosphate (g6p), by Merck Group (4 mentions)
Heparin, by Merck Group (4 mentions)
4 nitrocatechol, by Merck Group (4 mentions)
Testosterone, by Merck Group (4 mentions)
Glutathione (gsh), by Merck Group (4 mentions)
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Glucose 6 phosphate dehydrogenase, by Merck Group (3 mentions)
Cytochrome c, by Merck Group (3 mentions)
Theophylline, by Merck Group (3 mentions)
Benzyloxiresorufin, by Merck Group (2 mentions)
Nicotinamide adenine dinucleotide phosphate, by Merck Group (2 mentions)
21 protocols using methoxyresorufin
1
In Vitro Characterization of CYP1A1
2
Cytochrome c Enzymatic Assays in Cells
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Cytochrome c (cyt c) (horse heart), glucose-6-phosphate (G6P), glucose-6-phosphate dehydrogenase (G6PD), nicotinamide-adenine dinucleotide phosphate (NADPH), ethoxyresorufin, methoxyresorufin, resorufin, coumarin, 7-hydroxy coumarin, 3-cyano-7-ethoxycoumarin, 3-cyano-7-hydroxycoumarin, fluorescein, trypsin, penicillin-streptomycin (10,000 units penicillin and 10 mg streptomycin per mL), Dulbecco’s Modified Eagle’s Medium-low glucose (DMEM), fetal bovine serum (FBS), phosphate buffered saline pH 7.4 (PBS) and doxorubicin were obtained from Sigma Aldrich (St. Louis, MO, USA). Nicotinamide adenine dinucleotide phosphate (NADP+) was obtained from Gerbu (Heidelberg, Germany). Bradford reagent was obtained from Bio-Rad (Hercules, CA, USA) and Quiazol from Qiagen (Hilden, Germany). Dibenzylfluorescein, sodium dithionite, dimethyl sulfoxide (DMSO), acetonitrile (ACN) and sodium hydrogencarbonate (NaHCO3) were purchased from Merck (Kenilworth, NJ, USA). Insulin was obtained from Cell Applications, Inc. (San Diego, CA, USA). All other chemicals and solvents were of the highest grade commercially available.
3
Synthesis and Characterization of AαC Metabolites
4
Enzyme Assay for Cytochrome P450
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Methoxyresorufin, phenacetin, NADP+, and Glucose-6-phosphate were purchased from SigmaAldrich (St. Louis, MO, USA). Escherichia coli DH5α cells were purchased from Invitrogen (Carlsbad, CA, USA). Other chemicals were of the highest commercially available grade.
5
Cytotoxicity and Oxidative Stress Analysis
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Aflatoxin B1 (AFB1), sodium azide (NaN3), phenylmethylsulfonyl fluoride (PMSF), resorufin, ethoxyresorufin, methoxyresorufin, erythromycin, phorbol 12-myristate 13-acetate (PMA), dihydrorhodamine 123 (DHR), and lipopolysaccharides (LPS) were obtained from Sigma Aldrich Corp (St. Louis, MO, USA). 2-Amino-3,4 dimethylimidazo [4,5-f] quinolone (MeIQ), 2-aminoanthracene (2-AA), and 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide (AF-2) were provided by Wako Pure Chemicals (Osaka, Japan). Furthermore, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) were acquired from Merck (Darmstadt, Germany). Collagenase type IV and 4′-6-diamidino-2-phenylindole (DAPI) were bought from the Gibco/Invitrogen Corp (Waltham, MA, USA). Fetal bovine serum (FBS), Dulbecco’s Modified Eagle Medium (DMEM) and Roswell Park Memorial Institute (RPMI) medium were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). All standard phytochemicals were of high-performance liquid chromatography (HPLC) grade, and all other chemicals were of analytical grade.
6
Evaluation of Xylopic Acid's Effect on CYPs
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Phenobarbitone, ketoconazole, CYP450 isoenzymes, potassium phosphate dibasic, potassium phosphate monobasic, bovine serum albumin (BSA), nicotinamide adenine dinucleotide phosphate (NADPH), ethoxyresorufin, methoxyresorufin, pentoxyresorufin, benzyloxyresorufin, dextromethorphan, diclofenac, zinc sulphate heptahydrate, trimethylamine, and acetonitrile used to evaluate the effect of xylopic acid on CYPs were all obtained from Sigma-Aldrich Chemie GmbH (Eschenstrasse), Germany. XA was purified following the procedure described by Biney et al. [24 ].
7
Serum Enzyme Activity Assay in Rats
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The ALT and AST activity in serum of rats were determined using commercial kits produced by BioMar (Gliwice, Poland). Normal and pathological serum were used for quality control. Cytochrome P450 1A (CYP1A) subfamily in mammals consists of 2 isoforms, CYP1A1 and CYP1A2. Activity of the enzymes in liver of rats was measured using the kinetic method described by Dey et al. [26 (link)] and Zamaratskaia et al. [27 (link)] with ethoxyresorufin (CAS #5725-91-7, Sigma-Aldrich #46121-5MG-F, Saint Louis, USA) and methoxyresorufin (CAS #5725-89-3, Sigma-Aldrich #69125-5MG, Saint Louis, USA) as reagents. The fluorescence of both enzymatic reactions was measured at 535 nm and 590 nm as excitation/emission wave lengths, protein concentration was analyzed with Bradford reagent. Bovine serum albumin was used as standard.
8
Comprehensive Assay Protocol for Hepatic Function
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DAS, thioacetamide, ethoxyresorufin, methoxyresorufin, pentoxyresorufin, benzyloxyresorufin, resorufin, p-nitrophenol, erythromycin, glucose 6-phosphate, NADPH, glucose 6-phosphate dehydrogenase and the kits for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) assays were purchased from Sigma Chemical Company (St. Louis, MO, USA). The primary antibodies against individual CYP proteins were purchased from Easy-bio system (Seoul, Korea). Earle's balanced salt solution (EBSS) and guinea pig complements were purchased from GIBCO (Grand Island, NY, USA). Sheep red blood cells (SRBCs) were obtained at the College of Natural Resources at Yeungnam University. All other chemicals were of reagent grade commercially available and used as received.
9
Cytochrome P450 Enzyme Assay Protocol
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Beta-naphtoflavone (β-NF), 7-ethoxyresorufin (EROD), methoxyresorufin (MROD), benzyloxiresorufin (BROD), penthoxyresorufin (PROD), resorufin and benzo[a]pyrene (BP) were purchased from Sigma (St. Louis MO, USA). Culture media were obtained from BD Difco (Trenton, NJ, USA). The Escherichia coli DH5α recombinant and Salmonella typhimurium strains were kindly provided by Dr. Peter Guengerich (Vanderbilt University, Nashville, TN, USA).
10
Streptozotocin-Induced Diabetes Assay
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Streptozotocin (STZ), reduced and oxidized glutathione (GSH/GSSG), 1-chloro 2,4-dinitrobenzene (CDNB), cumene hydroperoxide, glutathione reductase, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), NADH, NADPH, cytochrome c, coenzyme Q2, sodium succinate, antimycin A, dodecyl maltoside, resorufin, 7-ethoxyresorufin, methoxyresorufin, Hoechst 33342, and ATP bioluminescent somatic cell assay kits were purchased from Sigma-Aldrich (St. Louis, MO, USA). 2′,7′-Dichlorofluorescein diacetate (DCFDA) was procured from molecular probes (Eugene, OR, USA). Kits for nitric oxide and caspase-3 and caspase-9 assays were purchased from R&D Systems Inc., MN, USA, and that for lipid peroxidation (LPO) from Oxis International Inc. (CA, USA). Kits for GSH/GSSG assay were procured from Promega Corp. (Madison, WI, USA). Apoptosis detection kits for flow cytometry were purchased from BD Pharmingen (BD Biosciences, San Jose, USA). Rin-5F cells were obtained from American Type Culture Collection (Manassas, VA, USA). Polyclonal antibodies against beta-actin, caspase-3, PARP, NOS-2, Nrf2, GLUT 2, Bax, Bcl-2, Akt, and p-Akt were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Reagents for cell culture, SDS-PAGE, and Western blot analyses were purchased from Gibco BRL (Grand Island, NY, USA) and Bio-Rad Laboratories (Richmond, CA, USA).
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