Acetonitrile acn

Bradykinin (SC090), culture and reagents were all purchased from Sigma-Aldrich (MO, St Louis, USA). DC Protein^TM assay, electrophoresis reagents and Clarity^TM Western ECL were purchased from Bio-Rad (Hercules, California, USA). PD 98,059 and Ibuprofen were ordered from Cayman Co (MI, Ann Arbor, USA). Antibodies used as follow: Anti-β-Actin (Abcam, Cambridge, UK, ab8227), Phospho-p44/42 MAPK (ERK1/2) rabbit mAb (Cell Signaling Technology, Danvers, MA, USA, 4370 s), p42/44 MAPK rabbit mAb (ERK1/2) (Cell Signaling Technology, 4695 s), Peroxidase-conjugated affinipure donkey anti-rabbit IgG (Jackson ImmunoResearch, West Grove, PA, USA) 711-035-152), or affinipure donkey anti-mouse IgG (Jackson Immune, 715–035-150), mouse monoclonal antibody anti-COX-2 (clone COX214). HPLC water was purchased from Avantor (Valley Center, PA, USA), acetonitrile (ACN) from Thermo Fisher Scientific (Waltham, MA, USA), trypsin/Lys-C mixture was produced by Promega (Madison, WI, United States) and formic acid (FA) was obtained from (TCI America, Portland, OR, USA).

Cytochrome c from equine heart (ref. C7752: MW~12.4 kDa, purity ≥ 95%), acetic acid (glacial), and Dulbecco’s Phosphate Buffered Saline (pH 7.4) were purchased from Sigma-Aldrich (Buchs, Switzerland). Sodium chloride and sodium acetate were obtained from Fisher Scientific (Loughborough, UK) and Fluka (Buchs, Switzerland), respectively.
Silver wire and silver chloride (AgCl) for the preparation of electrodes were obtained from Sigma Aldrich (Steinheim, Germany). PVC tubing (3 mm ID, 5 mm OD, and 1 mm wall) and agarose (used to prepare salt bridge assemblies) were obtained from VWR International AG (Dietikon, Switzerland) and Conda (Madrid, Spain), respectively.
The organic solvents, acetonitrile (ACN) and methanol (MeOH), were HPLC grade (Fischer Scientific, Loughborough, UK). Trifluoroacetic acid (TFA; 99% extra pure) was obtained from Acros organics (Geel, Belgium). Ultra-pure water (Millipore Milli-Q Gard 1 Purification pack resistivity > 18 MΩ.cm; Zug, Switzerland) was used to prepare all solutions. All other chemicals were at least of analytical grade.

LOX (2-(4-((2-Oxocyclopentyl)methyl)phenyl)propanoic acid, C₁₅H₁₈O₃, CAS ID 68767-14-5) was procured from Tokyo Chemicals Industry (Tokyo, Japan). LOX was used to synthesize cis-LOX and trans-LOX ((2S)-2-[4-[[(1R,2S)-2-hydroxycyclopentyl]methyl]phenyl]propanoic acid, C₁₅H₂₀O₃, CAS ID 83648-76-4) with purities of 96.5% and 97.9%, respectively [18 (link)]. DEX, KTC, DEXtromethorphan, and phenacetin were purchased from Sigma-Aldrich Co., LLC. (St. Louis, MO, USA) while midazolam was procured from Bukwang Pharmaceutical Co., Ltd. (Seoul, Korea). Mass spectrometry (MS) grade water and acetonitrile (ACN) were obtained from Fischer Scientific (Pittsburgh, PA, USA).

DMEM (Dulbecco’s modified eagle medium) powder was procured from Gibco by Life Technology (Thermo Fisher Scientific, Waltham, MA, USA). Sodium bicarbonate (NaHCO₃), calcium chloride (CaCl₂.2H₂O), and alpha-ketoglutaric acid disodium salt hydrate salts were purchased from Sigma-Aldrich (St Louis, MO, USA). Anti-cancer drug AZ628, DMSO (Dimethyl sulphoxide) and thiazolyl blue tetrazolium bromide (MTT) were obtained from Sigma–Aldrich (St Louis, MO, USA). DMEM-high glucose liquid media, FBS (Fetal Bovine Serum), TrypLE Express and penicillin-streptomycin were acquired from Sigma–Aldrich (St Louis, MO, USA). Acetonitrile (ACN) was from Fischer Scientific (Loughborough, UK). Bradford 1× dye reagent was purchased from Bio-rad Laboratories (Hercules, CA, USA).

Loxoprofen was purchased from Tokyo Chemicals Industry (Tokyo, Japan). Trans-OH and Cis-OH loxoprofen (purities 96.5% and 97.9%, respectively) were chemically synthesized from loxoprofen [22 (link)]. The β-reduced nicotinamide adenine dinucleotide phosphate (β-NADPH) regeneration system (NGS) was purchased from Promega Corp. (Madison, WI, USA). Pooled HLMs (mixed gender) were purchased from Sekisui Xeno Tech, LLC (Kansas City, MO, USA). Mixed Gender Corning UltraPool 150-donor liver cytosols; mouse, rat, dog, and monkey liver microsomes (RLM, MLM, DLM, and MoLM, respectively); and purified human recombinant cDNA encoding CYPs and UGTs were purchased from Corning Gentest (Woburn, MA, USA). Alamethicin and Uridine 5′-diphosphoglucuronic acid trisodium (UDPGA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Mass spectrometry (MS) grade water and acetonitrile (ACN) were acquired from Fischer Scientific (Pittsburgh, PA, USA).

Urea, 3-[(3-chloromidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium fluoride (NaF), sodium orthovanadate (Na₃VO₄), ethylene diamine tetraacetic acid (EDTA), phenylmethanesulfonyl fluoride (PMSF) and alpha-cyano-4-hydroxycinnamic acid (CHCA) were purchased from Sigma-Aldrich (St. Louis, MO). Trifluoroacetic acid (TFA) and acetonitrile (ACN) were obtained from Thermo Scientific (Rockford, IL).

MIBG, crizotinib, lorlatinib, and glyburide were purchased from Sigma-Aldrich (St. Louis, MO, USA). Erlotinib was kindly provided by Dr. Rheem Totah. Brigatinib was purchased from MedChemExpress (Monmouth Junction, NJ, USA). [¹⁴C]metformin (112 mCi/mmol) and [³H]1-methyl-4-phenylpyridinium (MPP+, 80 Ci/mmol) were purchased from American Radiolabeled Chemicals, Inc. (St. Louis, MO, USA). Acetonitrile (ACN) and formic acid (liquid chromatography with tandem mass spectrometry (LC-MS/MS)-grade) were obtained from Thermo Fisher Scientific (Rockford, IL, USA). Reagents for cell culture were purchased from Invitrogen (Carlsbad, CA, USA).