HDC (>98.0%) and H. helix extract were obtained from the Lab. of Pharmacognosy, College of Pharmacy, Yonsei University (Incheon, Korea). The extract was prepared by extracting the pulverized ivy leaves with 30% ethanol for 1 h using sonication. A voucher specimen of H. helix extract (HY-2016-01-05) was deposited at the Herbarium of the College of Pharmacy, Hanyang University, Ansan, Korea. H. helix extract contains 8.2% of HDC. The content of HDC was determined by liquid chromatography–tandem mass spectrometry (LC-MS/MS) [10 (link)] and the representative chromatogram is shown in Figure 4 . Pooled human liver microsomes and recombinant CYP2C8, CYP2C19, and CYP2D6 isozymes were purchased from BD Gentest (Woburn, MA, USA). Glucose-6-phosphate, β-NADP+, Glucose-6-phosphate dehydrogenase, coumarin, phenacetin, diclofenac, midazolam, mephenytoin, dextromethorphan, ketoconazole, and terfenadine were purchased from Sigma Chemical Co. (Saint Louis, MO, USA). All other solvents used were of HPLC grade and were obtained from J. T. Baker (Phillipsburg, NJ, USA). Distilled water was prepared using a Milli-Q purification system (Millipore, Billerica, MA, USA). All standard solutions and mobile phases were passed through a 0.22 µm membrane filter before use.
β nadp
Manufactured by Merck Group
Sourced in United States, Germany
β-NADP+ is a cofactor used in various enzymatic reactions. It functions as an electron acceptor in oxidation-reduction processes.
Automatically generated - may contain errors
Lab products found in correlation
Glucose 6 phosphate dehydrogenase, by Merck Group (9 mentions)
Glucose 6 phosphate (g6p), by Merck Group (8 mentions)
Phenacetin, by Merck Group (5 mentions)
Coumarin, by Merck Group (5 mentions)
Midazolam, by Merck Group (4 mentions)
Dextromethorphan, by Merck Group (4 mentions)
Diclofenac, by Merck Group (4 mentions)
Mephenytoin, by Merck Group (3 mentions)
Ketoconazole, by Merck Group (3 mentions)
Terfenadine, by Merck Group (3 mentions)
Milli q purification system, by Merck Group (3 mentions)
Pooled human liver microsomes, by BD (3 mentions)
Acetonitrile, by Merck Group (2 mentions)
Formic acid, by Merck Group (2 mentions)
Arachidonic acid, by Merck Group (1 mentions)
Reduced nicotinamide adenine dinucleotide, by Merck Group (1 mentions)
Nicotinamide adenine dinucleotide, by Merck Group (1 mentions)
Rhodamine 123, by Merck Group (1 mentions)
Adenosine diphosphate adp, by Merck Group (1 mentions)
Enzyme linked immunosorbent assay elisa kit, by Thermo Fisher Scientific (1 mentions)
13 protocols using β nadp
1
Quantitative Analysis of Hederagenin Derivative
2
Bicyclol Efficacy and Mechanism Study
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Bicyclol was kindly provided by the Beijing Union Pharmaceutical Plant (purity >99%). RIF and PZA were the products of J&K Scientific Ltd. (Beijing, China). INH, Rhodamine 123, palmitoyl-CoA, oxaloacetate, adenosine diphosphate (ADP), rotenone, antimycin A, ubiquinone-1, chlorzoxazone (CZX), 6-hydroxyl-chlorzoxazone (6-OH-CZX), arachidonic acid, cytochrome c, reduced nicotinamide adenine dinucleotide (NADH), glucose 6-phosphate, β-NADP, glucose 6-phosphate dehydrogenase, phenacetin, resveratrol, and nicotinamide adenine dinucleotide (NAD) were purchased from Sigma Chemical Co. (St Louis, MO, USA). Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) assay kits were obtained from BHKT Chemical Reagent Co., Ltd. (Beijing, China). TBIL, MDA, AKP, SOD, GSH, CAT and GSH-Px kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Enzyme-linked immunosorbent assay (ELISA) kits were products from eBioscience (San Diego, CA, USA). Other chemicals were of analytical grade and were obtained from the local market.
3
Vonoprazan Metabolism in Hepatic Microsomes
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Both vonoprazan acetate (purity 99.5%) and vonoprazan fumarate (purity 99.7%) and their metabolite M-I (purity 98.44%) were provided by Jiangsu Carefree Pharmaceutical Co. Ltd. Research Institute (Nanjing, China). The internal standard repaglinide (purity 99.7%) was purchased from National Institutes for Food and Drug Control (Beijing, China). Dog (male beagle dog, Lot. UKHU, mixed from 3 dogs) and human (male Mongolia human, Co. Ltd. SHQY, mixed from 11 individuals) hepatic microsomes were purchased from Research Institute of Liver Diseases (Shanghai, China). D-glucose-6-phosphate, glucose-6-phosphate dehydrogenase and β-NADP+ were purchased from Sigma-Aldrich (St. Louis, Mo, USA). Other chemicals and reagents were of analytical grade or better.
4
Metabolism and Kinetics Evaluation
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YZG-331, M2, and YZG-441 (purity ≥ 99%) were obtained from the Department of Natural Products Chemistry (Institute of Materia Medica, Chinese Academy of Medical Sciences). NADPH, NADH, β-NADP, glucose-6-phosphate (6-G-P), glucose-6-phosphate dehydrogenase (6-G-P-DH), midazolam, vanillin, 4-methylumbelliferyl-β-D -glucuronide (4MUG), benzydamine and aminobenzotriazole (ABT) were the products of Sigma Aldrich (St. Louis, MO, United States). Male mouse and rat liver microsomes (LMs)/cytosols were prepared by our group. Pooled mixed-gender human liver microsomes, male beagle dog/monkey liver microsomes, pooled mixed-gender dog/monkey/human liver cytosols, and recombinant human cytochrome P450 enzymes, recombinant human flavin-containing monooxygenases (FMOs) were obtained from BD Gentest (Woburn, MA, United States). HPLC-grade acetonitrile and methanol were purchased from Merck (Darmstadt, Germany). Deionized water was obtained from Milli-Qsystem (Millipore Co., Billerica, MA, United States). All other reagents and solvents were commercially available.
5
Cytochrome P450 Inhibition by E. longifolia
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The methanolic extract of E. longifolia roots was provided by the Institute of Marine Biochemistry, Vietnam Academy of Science and Technology (Hanoi, Vietnam). Pooled human liver microsomes and recombinant CYP1A2, CYP2A6, and CYP2C19 isozymes were purchased from BD Gentest (Woburn, MA, USA). Glucose-6-phosphate, β-NADP+, Glucose-6-phosphate dehydrogenase, phenacetin, coumarin, diclofenac, mephenytoin, dextromethorphan, midazolam, ketoconazole, and terfenadine were obtained from Sigma Chemical Co. (Saint Louis, MO, USA). All other solvents used were of HPLC grade and were purchased from J. T. Baker (Phillipsburg, NJ, USA). Distilled water was prepared using a Milli-Q purification system (Millipore, Billerica, MA, USA).
6
Investigating S. flavescens Root Extract Metabolism
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S. flavescens root extract was purchased from the Korea Plant Extract Bank (Chungbuk, Korea). The extract was prepared by extraction with a 70% ethanol solution. Glucose 6-phosphate, β-NADP+, glucose-6-phosphate dehydrogenase, phenacetin, coumarin, bupropion, diclofenac, S-mephenytoin, dextromethorphan, and chlorpropamide were purchased from Sigma Aldrich (St. Louis, MO, USA); rosiglitazone and midazolam from Toronto Research Chemicals (Toronto, ON, Canada); and kushenols A, C, I, and M, leachianone A, and sophoraflavone G from Core Sciences (Seoul, Korea). Pooled human liver microsomes, and baculovirus-insect cell expressed 2B6 and 3A4 were purchased from BD Gentest (Woburn, MA, USA). All other reagents were of the highest grade commercially available.
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S. flavescens root extract was purchased from the Korea Plant Extract Bank (Chungbuk, Korea). The extract was prepared by extraction with a 70% ethanol solution. Glucose 6-phosphate, β-NADP+, glucose-6-phosphate dehydrogenase, phenacetin, coumarin, bupropion, diclofenac, S-mephenytoin, dextromethorphan, and chlorpropamide were purchased from Sigma Aldrich (St. Louis, MO, USA); rosiglitazone and midazolam from Toronto Research Chemicals (Toronto, ON, Canada); and kushenols A, C, I, and M, leachianone A, and sophoraflavone G from Core Sciences (Seoul, Korea). Pooled human liver microsomes, and baculovirus-insect cell expressed 2B6 and 3A4 were purchased from BD Gentest (Woburn, MA, USA). All other reagents were of the highest grade commercially available.
7
Enzymatic NADP+ Quantification Assay
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NADPH was enzymatically converted to NADP+ following the procedure described by Vaseghi et al. (1999) (link), and the NADP+ concentration was assayed by the enzymatic cycling method (Vaseghi et al., 1999 (link)) with minor modifications. Briefly, a reaction mixture containing 0.1 M bicine, 0.1 M nicotinamide, 1 mM PES (phenazine ethosulfate), 4.2 mM MgSO4, 12.7 mM G6P (glucose 6-phosphate) and dinucleotide extracts in a final volume of 250 μL was prepared. The reaction was initiated by addition of glucose 6-phosphate dehydrogenase (G6PDH) and thiazoylyl blue tetrazolium bromide to final concentrations of 2.8 U and 0.25 mM, respectively. Water was used as blank and standard curves were obtained with β-NADP+ (Sigma). Reactions were monitored at 570 nm, for 30 min, using a Multiskan GO Microplate Spectrophotometer (Thermo Fisher Scientific).
8
Kinsenoside Purity and CYP Enzyme Assay
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Kinsenoside was contributed by Prof. Zhang, Tongji Medical College of Huazhong University of Science and Technology (Wuhan, China). Kinsenoside purity was determined by a high performance liquid chromatography with an evaporative light scattering detector (ELSD), which was >98% [37 (link)]. The HPLC-ELSD chromatogram of kinsenoside is shown in Figure 5 .
Pooled human liver microsomes and recombinant CYP2A6 were purchased from BD Gentest (Woburn, MA, USA). Glucose-6-phosphate, β-NADP+, Glucose-6-phosphate dehydrogenase, phenacetin, coumarin, diclofenac, mephenytoin, dextromethorphan, midazolam, ketoconazole, and terfenadine were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Amlodipine and lovastatin were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other solvents used were of HPLC grade and were purchased from J. T. Baker (Phillipsburg, NJ, USA). Distilled water was prepared using a Milli-Q purification system (Millipore, Billerica, MA, USA). All standard solutions and mobile phases were passed through a 0.22 µm membrane filter before use.
Pooled human liver microsomes and recombinant CYP2A6 were purchased from BD Gentest (Woburn, MA, USA). Glucose-6-phosphate, β-NADP+, Glucose-6-phosphate dehydrogenase, phenacetin, coumarin, diclofenac, mephenytoin, dextromethorphan, midazolam, ketoconazole, and terfenadine were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Amlodipine and lovastatin were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other solvents used were of HPLC grade and were purchased from J. T. Baker (Phillipsburg, NJ, USA). Distilled water was prepared using a Milli-Q purification system (Millipore, Billerica, MA, USA). All standard solutions and mobile phases were passed through a 0.22 µm membrane filter before use.
9
Amoxicillin Metabolism in Human Liver
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Amoxicillin (AMOX) was supplied by Sigma-Aldrich (Schnelldorf, Germany). Human liver microsomes (HLM), glucose-6-phosphate, β-NADP+, glucose-6-phosphate dehydrogenase and uridine 5′-diphosphoglucuronic acid triammonium salt (UDPGA) were obtained from Sigma-Aldrich (Schnelldorf, Germany). Other chemicals of analytical quality (acetonitrile, formic acid) were purchased from Merck (Darmstadt, Germany). Water was obtained by means of Milli-Q RG apparatus by Millipore (Millipore Intertech, Bedford, MA, USA) in our laboratory.
10
Cytochrome P450 Enzyme Assay Protocol
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Benzyloxy-, ethoxyresorufin, coumarin, EDTA, pyrazole, Bradford reagent, BSA, β-NADP, glucose-6-phosphate, glucose-6-phosphate dehydrogenase, resorufin, umbelliferone and glycine were from Sigma Chemical Co (St. Louis, MO, USA). Phenobarbital (Fenocris®) was from Cristália Produtos Químicos Farmacêuticos LTDA (São Paulo, Brazil). All other chemicals used in the experiments were of high analytical grade.
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