Licochalcone A (E-97%, Z-3% by NMR, Electronic Supplemental Material S1) and β-nicotinamide adenine dinucleotide phosphate, reduced sodium salt (NADPH), uridine 5’-diphosphoglucuronic acid triammonium salt (UDPGA), sulfatase, β-glucuronidase, sulfur trioxide-pyridine complex, and glutathione (GSH) were purchased from Sigma-Aldrich (St. Louis, MO). Pooled human liver microsomes (20 mg/mL, 150 men and women donors), cDNA-expressed human cytochrome P450 1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, and CYP3A4 (0.5 nmol of P450 in 0.5 mL) and cDNA-expressed recombinant human UGTs including UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B4, UGT2B7, UGT2B15, and UGT2B17 (5 mg protein/mL), were purchased from Corning Life Sciences (Tewksbury, MA). HPLC-grade solvents were purchased from Thermo Fisher (Pittsburgh, PA). Cryopreserved human hepatocytes (50 donors pooled) and manufacturer-specified cell culture media were purchased from Thermo Fisher (Waltham, MA).
Ugt1a4
Manufactured by Corning
UGT1A4 is a recombinant protein expressed in insect cells and purified for use in in vitro assays. It functions as a UDP-glucuronosyltransferase, an enzyme involved in the glucuronidation of various endogenous and exogenous compounds.
Automatically generated - may contain errors
Lab products found in correlation
Ugt1a9, by Corning (3 mentions)
Ugt2b7, by Corning (3 mentions)
Ugt1a3, by Corning (3 mentions)
Ugt2b15, by Corning (3 mentions)
Ugt1a6, by Corning (2 mentions)
Ugt1a1, by Corning (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Sulfur trioxide pyridine complex, by Merck Group (1 mentions)
Cyp2a6, by Corning (1 mentions)
Cyp2b6, by Corning (1 mentions)
Ugt2b4, by Corning (1 mentions)
Licochalcone a, by Merck Group (1 mentions)
Cyp2c8, by Corning (1 mentions)
Ugt1a10, by Corning (1 mentions)
Glutathione (gsh), by Merck Group (1 mentions)
β glucuronidase, by Merck Group (1 mentions)
Hplc grade solvents, by Thermo Fisher Scientific (1 mentions)
Potassium phosphate, by Merck Group (1 mentions)
Zidovudine, by Merck Group (1 mentions)
Raloxifene, by Merck Group (1 mentions)
3 protocols using ugt1a4
1
In Vitro Licochalcone A Metabolism
2
Palbociclib Glucuronidation and Sulfation Assay
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ABT (Sigma‐Aldrich), palbociclib sulfate (TRC), uridine 5′‐diphosphoglucuronic acid, UDPGA (Sigma‐Aldrich), MgCl2 (Sigma‐Aldrich), Alamethicin (Sigma‐Aldrich), dimethyl sulfoxide, DMSO (Sigma‐Aldrich), ACN (liquid chromatography‐mass spectrometry [LC–MS] grade; Fisher Chemical), ACN (Optima LC/MS grade, Fisher Chemical) formic acid (Fisher Chemical), potassium phosphate (Sigma‐Aldrich) 5,5‐diethyl‐1,3‐diphenyl‐2‐iminobarbituric acid (“39:an”; Sigma‐Aldrich), D‐Saccharolactone (Sigma‐Aldrich), ultra‐pure water (Millipore), recombinant UGT supersomes (UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, UGT2B7, and UGT2B15; Corning), β‐estradiol (Sigma‐Aldrich), chenodeoxycholic acid (CDCA; Sigma‐Aldrich), trifluoperazine (TFP; Sigma‐Aldrich), serotonin (Sigma‐Aldrich), propofol (Sigma‐Aldrich), zidovudine (Sigma‐Aldrich), oxazepam (Sigma‐Aldrich), raloxifene (Sigma‐Aldrich), dehydroepiandrosterone (DHEA; Sigma‐Aldrich), DHEA‐sulfate (Sigma‐Aldrich), 4‐methylumbelliferone (4‐MU; Sigma‐Aldrich), palbociclib sulfate (Toronto Research Chemicals); recombinant sulfotransferases (SULT1A1, SULT1A3, SULT1B1, SULT1C2, SULT1C4, SULT1E1, SULT2A1, and SULT2B1; R&D Systems), palbociclib, and ARV‐471 were synthesized by AZ chemistry.
3
In Vitro Metabolism of Bempedoic Acid and ESP15228
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The in vitro metabolism of [ 14 C] bempedoic acid (60 mM) and [ 14 C] ESP15228 (8 mM) was studied in cDNA-expressed recombinant uridine 5 0diphospho-glucuronosyltransferase (UGT; 1.0 mg/ml protein) enzyme preparations in the presence of 2 mM UDPGA at 37 C. Recombinant UGT enzymes (Supersomes) prepared from baculovirus-infected insect cells were procured from Corning Inc. (Woburn, MA): UGT1A1 (Lot 3273576), UGT1A3 (Lot 3296661), UGT1A4 (Lot 3303828), UGT1A6 (Lot 4036003), UGT1A9 (Lot 3287625), UGT2B7 (Lot 3287588), and UGT2B15 (Lot 3247829). Incubation reactions with each recombinant UGT enzyme were conducted in triplicate and terminated after 0 and 60 minutes with addition of ice-cold acetonitrile containing 6% acetic acid v/v (1 ml). Samples were vortex mixed and centrifuged with transfer of the resultant supernatants to a clean tube and stored at À20 C until analysis by HPLC with radiometric detection. Individual UGT enzyme (0.5 mg/ml protein) activities were confirmed using 4-methylumbelliferone (100 mM) as a positive control. The rates of 4-methylumbelliferone glucuronidation were $1.43 nmol/min/mg protein (range, 1.43-19.8 nmol/min/mg protein) across UGT enzyme preparations.
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