Gentest atpase assay kit

Ferulic acid, sulforhodamine B (SRB), (+/−)-verapamil hydrochloride and other reagents for synthesis were purchased from Sigma-Aldrich (St. Louis, MO, USA). [³H]-daumnomycin (16 Ci/mmol) was supplied by PerkinElmer Life and Analytical Sciences (Boston, MA, USA). The BD Gentest^TM ATPase assay kit and human P-glycoprotein membranes were commercially provided from BD Bioscience (Woburn, MA, USA). Paclitaxel (PTX) was obtained from Samyang Genex (Daejeon, Korea). All reagents and solvents were supplied as synthesis grades, cell culture grades and HPLC analysis grades. Melting points were measured on an electro-thermal digital melting point (Buchi, Essen, Germany) without calibration. ¹H NMR spectra were recorded on Varian NMR AS and Varian Unity Inova 400 MHz NMR spectrometers. Chemical shifts were reported in parts per million (d) units relative to the solvent peak. The ¹H NMR data were reported as peak multiplicities: s for singlet; d for doublet; t for triplet; and m for multiplet. Coupling constants were recorded in hertz. Mass spectra data was obtained on an Agilent 6220 Accurate-Mass time-of-flight liquid chromatography/mass spectrometry (TOF LC/MS). All the synthesized compounds have been registered as a patent (KR10-2013-0099676) by Prof. Hea-Young Park Choo and Ms So Yeon Lim since 2013.

The chemical markers (purity ≥98%) were purchased from Sichuan Victory Biological Technology Co., Ltd. Materials for tissue culture were purchased from Invitrogen (Carlsbad, CA). The antibodies were obtained from Abcam (Cambridge, United Kingdom), Absin (Absin Bioscience Co., Ltd.) and Invitrogen. BD GentestTM ATPase assay kit was purchased from BD Bioscience (San Jose, CA). GST activity assay kit, BCA protein assay kit, A549 cell lines were purchased from Dingguo Changsheng Biotechnology Ltd. (Guangzhou, China). A549-DDP (cisplatin resistance) cell was purchased from Fudan IBS cell bank (FDCC). TRIzol^™ Reagent, High-Capacity cDNA Reverse Transcription Kits, PowerUp^™ SYBR^™ Green Master Mix, Pierce^® Fast Western Blot Kit, and PVDF Transfer Membrane were purchased from ThermoFisher Scientific (Guangzhou, China). Materials that not specified here were purchased from Sigma-Aldrich. (St. Louis, MO), ThermoFisher Scientific, Merck (Whitehouse Station, NJ), Dingguo and Xinjing Biotechnology.

The activities of MRP 1, MRP 2, MRP 3, BCRP, and MDR 1/P-gp were assayed using the BD Gentest ATPase Assay kit (BD Biosciences, Woburn, MA) as described by the manufacturer.