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Pentoxyresorufin

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Pentoxyresorufin is a fluorescent dye used as a substrate in biochemical assays. It is a derivative of resorufin, a common fluorescent indicator. Pentoxyresorufin can be used to measure enzyme activity, particularly the activity of cytochrome P450 enzymes.

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Lab products found in correlation

Ethoxyresorufin, by Merck Group (7 mentions) Methoxyresorufin, by Merck Group (5 mentions) Diclofenac, by Merck Group (4 mentions) Nadph, by Merck Group (3 mentions) Benzyloxyresorufin, by Merck Group (3 mentions) P nitrophenol, by Merck Group (3 mentions) Resorufin, by Merck Group (3 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Dextromethorphan, by Merck Group (2 mentions) Testosterone, by Merck Group (2 mentions) Glutathione (gsh), by Merck Group (2 mentions) 1 chloro 2 4 dinitrobenzene, by Merck Group (2 mentions) 2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid abts, by Merck Group (1 mentions) Giemsa stain, by Merck Group (1 mentions) Analytical grade reagents, by Merck Group (1 mentions) 7 ethoxyresorufin, by Merck Group (1 mentions) Thiobarbituric acid tba, by Merck Group (1 mentions) Benzyloxiresorufin, by Merck Group (1 mentions) Supersomes, by BD (1 mentions) Potassium phosphate dibasic, by Merck Group (1 mentions)

Close spelling variants of this product

7-pentoxyresorufin (7 citations)

10 protocols using pentoxyresorufin

1

In Vitro Characterization of CYP1A1

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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).
Rodeiro I., Olguín S., Santes R., Herrera J.A., Pérez C.L., Mangas R., Hernández Y., Fernández G., Hernández I., Hernández-Ojeda S., Camacho-Carranza R., Valencia-Olvera A, & Espinosa-Aguirre J.J. (2015). Gas Chromatography-Mass Spectrometry Analysis of Ulva fasciata (Green Seaweed) Extract and Evaluation of Its Cytoprotective and Antigenotoxic Effects. Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 520598.
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2

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 ].
Agbenyeku M.A., Appiah-Opong R., Obese E., Biney R.P., Adakudugu E.A., Forkuo A.D., Osei S.A., Abeka M.K, & Ameyaw E.O. (2022). In Vitro and In Vivo Effect of Xylopic Acid on Cytochrome P450 Enzymes. Advances in Pharmacological and Pharmaceutical Sciences, 2022, 4524877.
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3

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.
Kim N.H., Lee S., Kang M.J., Jeong H.G., Kang W, & Jeong T.C. (2014). Protective Effects of Diallyl Sulfide against Thioacetamide-Induced Toxicity: A Possible Role of Cytochrome P450 2E1. Biomolecules & Therapeutics, 22(2), 149-154.
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4

Juvenile Rat Liver Micronucleus Assay

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Resorufin, ethoxyResorufin and pentoxyResorufin were purchased from Sigma-Aldrich Inc. (St. Louis, MO, U.S.A.). Tolbutamide, hydroxyl- tolbutamide, midazolam, 4-hydroxymidazolam, bufuralol and 1’-hydroxybufuralol were purchased
from Daiichi Chemical (Tokyo, Japan). Aniline and p-aminophenol were obtained from Wako Pure Chemical (Osaka, Japan). All the other chemicals were of analytical or HPLC grade.
Diethylnitrosamine (DEN) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Prior to administration, DEN was dissolved in physiological saline (Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan) and
administered by oral gavage. The dose for DEN was selected based on data that showed positive results in the juvenile rat liver MN assay [29 (link),30 (link),31 (link),32 (link),33 (link)]. Negative control animals received physiological saline (DEN solvent).
NAGASUE R., MURATA I., SASAKI K., SAKAI R., MIYAJIMA H, & SHIMODA M. (2017). Effectiveness of the liver micronucleus assay using juvenile mice. The Journal of Veterinary Medical Science, 79(7), 1310-1317.
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5

CYP450 Enzyme Activity Evaluation

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Gemfibrozil was obtained from Hunan Qianjin Xiangjiang Pharmaceutical Co. Ltd (Zhuzhou, China). Trizol was obtained from Invitrogen (CA, USA). The reverse transcription kit was purchased from Thermo Fisher (PA, USA). The LightCycle 480 SYBR Green I Master Mix was purchased from Roche. Antibodies against CYP3A (sc-30621) and CYP2B1/2 (sc-73546) were products by Santa Cruz Biotechnologies (CA, USA). Antibody against GAPDH (ab181602), CYP2C (ab137015) were obtained from Abcam (Cambridge, UK). Midazolam, testosterone, pentoxyresorufin, diclofenac were purchased from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA). 1′-hydroxy-Midazolam (1-OH-M), 6β-hydroxyl-testosterone (6β-OH-T), resorufin (Reso), and 4-hydroxy-diclofenac (4-OH-D) were obtained from Toronto Research Chemical Inc. (Toronto, ON, Canada). Purified water was freshly prepared using a Millipore Elix (Bedford, USA). All the other chemicals were of the highest grade from commercial sources.
Shi C., Min L., Yang J., Dai M., Song D., Hua H., Xu G., Gonzalez F.J, & Liu A. (2017). PPARα Activation Suppresses Cytochrome P450 Induction Potential in Mice Treated with Gemfibrozil. Basic & clinical pharmacology & toxicology, 121(3), 169-174.
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6

Comprehensive Enzymatic Assay Protocol

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Acetaminophen, bufurolol, 1’-hydroxybufurolol, caffeine, dexamethasone, ethoxyresorufin, glutathione (reduced), mephenytoin, 4-hydroxymephenytoin, pentoxyresorufin, resorufin, phenacetin, warfarin, 7-hydroxywarfarin, phenylmethanesulfonyl fluoride, p-nitrophenol (PNP), UDP-glucuronic acid (ammonium salt) (UDPGA), 1-chloro-2, 4-dinitrobenzene (CDNB), 2-mercaptoethanol, 3’-Phosphoadenosine-5’-phosphosulfate (PAPS), flavin adenine dinucleotide, dicoumarol, 2,6 dichlorophenolindophenol, 2-naphthylsulfate, high performance liquid chromatography (HPLC) grade acetonitrile, and methanol were purchased from Sigma–Aldrich (St. Louis, MI, USA). Nicotinamide adenine dinucleotide phosphate (NADPH) and dimethyl sulfoxide were purchased from SRL Pvt. Ltd (Mumbai, Maharashtra, India). Testosterone, 6b-hydroxyTestosterone, chlorzoxazone, and 6-hydroxychlorzoxazone were purchased from Cayman chemical company (USA). Ultrapure water (18.2 M/Ω cm) was obtained from Milli-Q PLUS PF water. All other chemicals were commercially available or HPLC grade.
Kumar D., Trivedi N, & Dixit R.K. (2016). Evaluation of the synergistic effect of Allium sativum, Eugenia jambolana, Momordica charantia, Ocimum sanctum, and Psidium guajava on hepatic and intestinal drug metabolizing enzymes in rats. Journal of Intercultural Ethnopharmacology, 5(4), 372-382.
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7

Assessing CYP450 Enzyme Activities

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Ethoxyresorufin-O-deethylase (EROD), methoxyresorufin-O-deethylase (MROD), and pentoxyresorufin-O-deethylase (PROD) assays were performed to determine the activities of CYP1A1, CYP1A2, and CYP2B as described previously [21 (link)]. The differentiated cells were treated with 1 μM 7-ethoxyresorufin (Fanbo biochemical, Beijing, China), methoxyresorufin (Fanbo biochemical), and pentoxyresorufin (Sigma-Aldrich) for 24 hours, respectively. The fluorescent products of CYP450 substrates leaked from cells were determined at a wavelength of 585 nM in the absorption under Infnite® 200 PRO (Tecan, Männedorf, Switzerland). Quantitative studies employed the high-purity reference standard of resorufin (Fanbo biochemical) for assay standardization.
Yuan J., Li W., Huang J., Guo X., Li X., Lu X., Huang X, & Zhang H. (2015). Transplantation of human adipose stem cell-derived hepatocyte-like cells with restricted localization to liver using acellular amniotic membrane. Stem Cell Research & Therapy, 6, 217.
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8

Ethical Evaluation of Chemical Compounds

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Ethoxyresorufin, pentoxyresorufin, diclofenac and dextromethorphan were purchased from Sigma-Aldrich (St Louis, MO, USA). All other chemicals and reagents used were of analytical grade and obtained from standard suppliers.
Ethical clearance for the study was obtained from the Institutional Review Board of the Noguchi Memorial Institute for Medical Research (NMIMR-IRB CPN 020/11-12), University of Ghana, Legon, Ghana.
Appiah-Opong R., Tuffour I., Ofori-Attah E., Aning A., Atchoglo P., Danso E.A., Ahedor B., Adjei S, & Nyarko A.K. (2018). In Vivo Modulation of Rat Liver Microsomal Cytochrome P450 Activity by Antimalarial, Anti-HIV, and Antituberculosis Plant Medicines. Journal of Evidence-based Integrative Medicine, 23, 2515690X18810001.
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9

Cytochrome P450 Activity Assays

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Bovine serum albumin (BSA), NADPH, 1-chloro-2,4-dinitrobenzene (CNDB), glutathione, o-phenylenediamine dihydrochloride (OPD), ethoxyresorufin, methoxyresorufin, benzyloxyresorufin and pentoxyresorufin, were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO). Microtiter plates (MaxiSorp) were purchased from Nunc (Roskilde, Denmark). Polyclonal CYP1A1 antibody was purchased from Biosense Laboratories, (Bergen, Norway). CYP2B6, CYP2E1 and CYP2B1/2 antibodies were purchased from Santa Cruz Biotechnology (Dallas TX, USA). The CYP3A antiserum was a kind gift from Dr. Malin Celander. All other chemicals were of the highest commercially available grade.
Arukwe A., Røsbak R., Adeogun A.O., Langberg H.A., Venter A., Myburgh J., Botha C., Benedetti M, & Regoli F. (2015). Biotransformation and Oxidative Stress Responses in Captive Nile Crocodile (Crocodylus niloticus) Exposed to Organic Contaminants from the Natural Environment in South Africa. PLoS ONE, 10(6), e0130002.
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10

Cytochrome P450 Enzyme Assay Protocol

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Testosterone, ethoxyresorufin, methoxyresorufin, pentoxyresorufin, resorufin, p-nitrophenol, 4-nitrocatechol, NADPH, glutathione, 1-chloro-2,4-dinitrobenzene, lauric acid, 12-hydroxy lauric acid, diclofenac (sodium salt), chlorzoxazone, dextromethophen, 1,1,3,3-tetraethoxypropan, thiobarbituric acid, and heparin were obtained from Sigma (St. Louis, MO, USA). 6-β-HydroxyTestosterone was purchased from Ultrafine Chemicals (Manchester, UK). All other chemicals and reagents were of analytical grade and were obtained commercially.
Yao H.T., Yang Y.H, & Li M.L. (2019). Intake of Molecular Hydrogen in Drinking Water Increases Membrane Transporters, p-Glycoprotein, and Multidrug Resistance-Associated Protein 2 without Affecting Xenobiotic-Metabolizing Enzymes in Rat Liver. Molecules, 24(14), 2627.
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