Glucose 6 phosphate dehydrogenase

Manufactured by Corning

Sourced in United States

Glucose-6-phosphate dehydrogenase is an enzyme that catalyzes the conversion of glucose-6-phosphate to 6-phosphoglucono-delta-lactone, the first step in the pentose phosphate pathway. It is an important component in many biochemical assays and laboratory procedures.

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3 protocols using glucose 6 phosphate dehydrogenase

Quantification of Deuterium-Labeled Ibrutinib and Metabolites

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Ibrutinib (free base; I‐3311) was purchased from LC Laboratories. Deuterium‐labeled ibrutinib ([²H₅]ibrutinib, d₅‐ibrutinib, 22,561) and PCI 45227 (M37) were purchased from Cayman Chemical. Midazolam (M‐908), 1′‐hydroxymidazolam (H‐902), and d₄‐1′‐hydroxymidazolam (H‐921) were purchased from Cerilliant Corporation. The 4β‐HC (700036) and cholesterol (700000P) were purchased from Avanti Polar Lipids. Deuterium‐labeled 4β‐HC ([²H₇]4β‐HC, d₇‐4β‐HC, H917982) and deuterium‐labeled cholesterol ([²H₇]cholesterol, d₇‐cholesterol, C432503) were purchased from Toronto Research Chemicals. Dimethyl sulfoxide (DMSO), Optima liquid chromatography mass spectrometry LC–MS grade water, Optima LC–MS grade acetonitrile, Optima LC–MS grade methanol were obtained from Fisher Scientific. All other chemicals and reagents were of analytical grade or higher.
As described previously,²³ an NADPH‐regenerating system, consisting of solution A (26 mM NADP⁺, 66 mM glucose 6‐phosphate, 66 mM MgCl₂ in water) and solution B (40 U/ml glucose 6‐phosphate dehydrogenase in 5 mM sodium citrate), was purchased from Corning Life Sciences. InVitroGRO Krebs–Henseleit buffer (KHB) medium (product no. Z99074) and thawing (HT) medium (product no. 990006) were purchased from BioIVT.

Lee J., Fallon J.K., Smith P.C, & Jackson K.D. (2022). Formation of CYP3A‐specific metabolites of ibrutinib in vitro is correlated with hepatic CYP3A activity and 4β‐hydroxycholesterol/cholesterol ratio. Clinical and Translational Science, 16(2), 279-291.

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In Vitro Metabolic Enzyme Assay

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Methanol, acetonitrile, ammonium formate, glucose-6-phosphate, β-nicotinamide adenine dinucleotide phosphate hydrate (NADP⁺), uridine 5′-diphosphoglucuronic acid trisodium salt (UDPGA), adenosine 3′-phosphate 5′-phosphosulfate triethylammonium salt (PAPS), S-(5′-adenosyl)-L-methionine p-toluenesulfonate salt (SAM), magnesium chloride hexahydrate and sodium citrate tribasic dihydrate were purchased from Sigma-Aldrich (St. Louis, MI, USA). Formic acid was acquired from Scharlab (Sentmenat, Barcelona, Spain) and Gentest™ 0.5 M phosphate buffer pH 7.4 and glucose-6-phosphate dehydrogenase from Corning (Woburn, MA, USA). Ultrapure water was obtained using a Mili-Q RG system from the company Milipore (Burlington, MA, USA). NEP hydrochloride reference material (98% purity) was purchased from Cayman Chemical (Ann Arbor, MI, USA) and a stock solution prepared at 1 mg/mL (4.1 mM free base) in Methanol. NEP working solutions of 40 and 732 μM were also prepared by appropriate dilution of NEP stock solution in Methanol. Dapaconazole, the positive control for the experiment, was supplied by Biolab Farmacêutica Ltda. (São Paulo, Brazil) and a stock solution prepared at 1 mg/mL (2.4 mM) in Methanol. A dapaconazole working solution of 40 μM was prepared from a dapaconazol stock solution in Methanol.

Godoi A.B., Antunes N.D., Cunha K.F., Martins A.F., Huestis M.A, & Costa J.L. (2024). Metabolic Stability and Metabolite Identification of N-Ethyl Pentedrone Using Rat, Mouse and Human Liver Microsomes. Pharmaceutics, 16(2), 257.

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Characterization of Drug Metabolism Assays

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Uridine 59-diphosphoglucuronic acid trisodium salt (UDPGA), NADPH, alamethicin from Trichoderma viride, adenosine 3-phosphate 5-phosphosulfate, diclofenac sodium salt, midazolam hydrochloride, felodipine, S-(+)-mephenytoin, raloxifene hydrochloride, lovastatin, nisoldipine, nifedipine, methadone, quinidine, testosterone, benzydamine hydrochloride, cisapride, cyclosporin, terbutaline, sildenafil, verapamil, trazodone, atorvastatin, simvastatin, buspirone, rifabutin, saquinavir, terfenadine, zolpidem, repaglinide, indinavir, alprazolam, carbazeran, enalapril, ramipril , and dabigatran etexilate were purchased from Sigma-Aldrich (St. Louis, MO). NADPH-regenerating system containing NADP + , glucose-6phosphate, and glucose-6-phosphate dehydrogenase was purchased from Corning (Wiesbaden, Germany). Organic solvents were of LC-mass spectrometry (MS) or higher quality grade and were acquired from VWR International (Radnor, PA) or Fisher Scientific UK Ltd (Loughborough, UK). Purified water was obtained from Milli-Q Integral 5 Water Purification System (Merck KGaA, Darmstadt, Germany). Phosphate buffer, HQM Hepatocyte/Enterocyte Incubation Medium, and Co-Factor N for MetMax were purchased from In Vitro ADMET Laboratories (Columbia, MD).

Davies M., Peramuhendige P., King L., Golding M., Kotian A., Penney M., Shah S, & Manevski N. (2020). Evaluation of In Vitro Models for Assessment of Human Intestinal Metabolism in Drug Discovery. Drug metabolism and disposition: the biological fate of chemicals, 48(11).

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