High pressure liquid chromatography grade solvents

The FinePure Universal DNA Purification Kit was purchased from GENFINE (Beijing, China). The CYP2J2 Human Tagged ORF Clone (RC207417) was purchased from Origene (United States). Saccharomyces cerevisiae strain YPH499 was obtained from ATCC (VA, United States). The yeast expression vector pESC-URA was purchased from Stratagene (United States). The following primary antibodies were used: rabbit polyclonal anti-CYP2J2 antibody (Abcam, United Kingdom), mouse monoclonal anti-OR antibody (Santa Cruz Biotechnology, United States) and corresponding secondary antibodies (Proteintech, China). Ebastine and terfenadine were purchased from Tokyo Chemical Industry. HydroxyEbastine, terfenadine alcohol and midazolam were purchased from Santa Cruz and TRC. The NADPH-regenerating system was purchased from Promega (United States). High-pressure liquid chromatography-grade solvents were purchased from Fisher Scientific Co., (United States). All the other chemicals and solvents were commercially available at analytical grade or the highest grade.

Materials and kits were purchased from the following sources: TIANamp Blood DNA Midi Kit (TIANGEN, Beijing, China); PrimeSTAR HS DNA polymerase and restriction enzymes (Takara Bio, Inc.; Otsu, Shiga, Japan); Spodoptera frugiperda (Sf)21 insect cells, Sf-900TM III SFM insect culture medium and Bac to-Bac Baculovirus Expression System (Invitrogen, Carlsbad, CA, United States); Mouse monoclonal anti-OR antibody (Santa Cruz Biotechnology, Dallas, Texas, United States); Rabbit polyclonal anti-CYP2C9 antibody (Abcam, Cambridge, United Kingdom); Super Signal West Pico Trial Kit (Thermo Scientific, Rockford, IL, United States); 4-Hydroxytolbutamide, 4-hydroxydiclofenac, and telmisartan (Toronto Research Chemicals, Inc.; Toronto, Ontario, Canada); Diclofenac and chlorpropamide (Tokyo Chemical Industry Co., Ltd.; Tokyo, Japan); Tolbutamide, losartan and E-3174 (Sigma-Aldrich, St. Louis, MO, United States); NADPH-regenerating system (Promega, Madison, WI, United States); High-pressure liquid chromatography-grade solvents (Fisher Scientific Co.; Fair Lawn, NJ, United States). All other chemicals and reagents were of analytical grade or the highest commercially available quality.