The authentic standards of ellagic acid (EA), pyrogallol, and gallic acid (GA) were purchased from the Chengdu MUST Bio-Technology CO. Ltd. (Chengdu, China), phosphate-buffered saline (PBS, pH = 7.4, 0.01 M), HPD resin was purchased from Tianjin Baowen Chemical Technology CO. Ltd. (Tianjin, China), anaerobic bags (AnaeroPouch Anaero, Mitsubishi Gas Chemical Company Inc., Tokyo, Japan), GAM broth medium was bought from Qingdao Haibo Biology CO. Ltd. (Qingdao, China), β-nicotinamide adenine dinucleotide phosphate (NADP), glucose-6-phosphate (G-6-P), glucose-6-phosphate dehydrogenase (G-6-PD), dithiothreitol, and S-adenosyl-L-methionine (SAM) were obtained from Sigma Chemical CO. (St. Louis, MO, USA). Acetonitrile, methanol, and formic acid were of HPLC-grade, which were purchased from Thermo Fisher Scientific CO. Ltd. (Waltham, MA, USA). Purified water was prepared by using Milli-Q System (Millipore, Billerica, MA, USA). All other analytical-grade reagents were provided by the Sinopharm Chemical Reagent CO. Ltd. (Shanghai, China).
Glucose 6 phosphate dehydrogenase g6pd
Manufactured by Merck Group
Sourced in United States
Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme that catalyzes the first step in the pentose phosphate pathway, converting glucose-6-phosphate into 6-phosphoglucono-δ-lactone and reducing NADP+ to NADPH. This enzyme plays a crucial role in maintaining the level of NADPH, an important cofactor for various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Glucose 6 phosphate (g6p), by Merck Group (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (2 mentions)
Milli q system, by Merck Group (1 mentions)
Formic acid, by Thermo Fisher Scientific (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
S adenosyl l methionine sam, by Merck Group (1 mentions)
β nicotinamide adenine dinucleotide phosphate nadp, by Merck Group (1 mentions)
Anaero pouch anaero, by Mitsubishi (1 mentions)
High performance liquid chromatography (hplc), by Thermo Fisher Scientific (1 mentions)
Tween 20, by Merck Group (1 mentions)
Quercetin, by Merck Group (1 mentions)
Digitonin, by Merck Group (1 mentions)
Cell culture media and supplements, by Thermo Fisher Scientific (1 mentions)
Feso4, by Merck Group (1 mentions)
Trizma base, by Merck Group (1 mentions)
Menadione, by Merck Group (1 mentions)
Esterase, by Merck Group (1 mentions)
6 protocols using glucose 6 phosphate dehydrogenase g6pd
1
Quantification of Antioxidant Compounds
2
Oxidative Stress Assay Protocol
3
Enzymatic Assay for LQ Biotransformation
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LQ (CAS No: 551-15-5) was purchased from Chengdu Desert Biotechnology Co., Ltd. (Chengdu, China). β-NADP (β-nicotinamine adenine dinucleotide phosphate), glucose-6-phosphate (G-6-P) and glucose-6-phosphatedehydrogenase (G-6-PD) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). MgCl2, UDPGA (uridine-5′-diphosphoglucuronic acid trisodium salt), Tris–HCl and alamethicin were purchased from BD Biosciences (Woburn, MA, USA). Phosphate-buffered saline (PBS) was purchased from Sangon Biotech Co., Ltd. (Shanghai, China). HPLC-grade methanol was purchased from J.T. Baker Chemical Company (Phillipsburg, NJ, USA). Formic acid (HPLC grade) was provided by Diamond Technology (Dikma Technologies Inc., Lake Forest, CA, USA). Purified water was obtained from Hangzhou Wahaha Group Co., Ltd. (Hangzhou, China).
4
Liver Microsomal Metabolism Evaluation
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Rat liver microsomes and human liver microsomes were purchased from Sekisui XenTech Co. (Kansas City, KS, United States). Human plasma was obtained from Rild Research Institute for Liver Disease Co. (Shanghai, China). Glucose-6-phosphate dehydrogenase (G-6-PD) and NADP were purchased from Sigma-Aldrich Co. (St. Louis, Mo, United States). Dulbecco’s modified eagle’s medium (DMEM), trypsogen and fetal bovine serum (FBS) were purchased from Invitrogen Co. (Grand Island, NY, United States). D-glucose-6-phosphate sodium salt (G-6-P) was purchased from Aladdin Industrial Co. (Shanghai, China). Dimethyl sulfoxide (DMSO) and methanol (chromatographic grade) were purchased from Sigma-Aldrich Co. (St. Louis, Mo, United States). Acetonitrile (chromatographic grade) was purchased from Thermo Fisher Scientific Co. (Waltham, MA, United States). The other chemicals used in this study were of analytical grade, and purchased from Sinopharm Chemical Reagent Co. (Shanghai, China).
5
Bilirubin Formation Assay Protocol
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This method was carried out as described previously with modifications35 (link). The treated cells were resuspended in homogenization buffer containing 0.25 M sucrose (Sigma, St. Louis, MO), 20 mM Tris–HCl at pH 7.4 (Bio-Rad Laboratories, Hercules, CA), 1 mM EDTA (Sigma, St. Louis, MO), and 0.1% protease inhibitor cocktail (Roche, Mannheim, Germany). Reaction mixture consisting of 100 μl sample, 2 mg/ml biliverdin reductase from rat liver, 1 mM β-nicotinamide adenine dinucleotide phosphate (β-NADPH; Sigma, St. Louis, MO), 2 mM glucose-6-phosphate (Sigma, St. Louis, MO), 1 U glucose-6-phosphate dehydrogenase (G6PD; Sigma, St. Louis, MO) and 25 μM hemin (Sigma, St. Louis, MO) was prepared fresh and incubated at 37 °C for 1 h in the dark. The formed bilirubin was extracted with chloroform and the change in optical density at 464–530 nm was measured in a microplate reader (Multiskan Go; Thermo Fisher Scientific, Waltham, MA). HO activity was expressed as picomoles of bilirubin formed per milligram of protein per hour by using an extinction coefficient of 40 mM-1/cm for bilirubin in chloroform and protein concentration of the sample.
6
Intracellular NADPH and NADP+ Levels
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NADPH activity was testedby a previously described method [45] (link). Following treatment, intracellular NADPH and NADP + levels were examined [46] . Briefly, two million cells were plated onto 10cm dishes and lysed in 300 μL of extraction buffer after the applied treatment [27] . For NADPH extraction, 100 μL of the supernatant was incubated at 60 °C for 30 min. Next, 100 μL of NADP-cycling buffer [27] with 1.0 U glucose-6-phosphate dehydrogenase (G6PD; Sigma-Aldrich) was added. After 1 min of incubation at 30 °C, 20 μL of 10 mM glucose 6-phosphate (G6P; Sigma-Aldrich) was added to the mixture, and the change in absorbance at 570 nm was measured. The concentration of NADP + was calculated by subtracting [NADPH] from [total NADP] [27] . The relative NADPH activity was calculated by determining the NADPH/NADP + ratio.
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