P-Glycoprotein / MDR1 (human) Drug Interaction Assay

The materials provided in the assay kit for MDR1 / Human P-Glycoprotein Drug Interaction are formatted to allow for two independent 48-well assay setups. Each aliquot of HCT-Pgp Cells is provided as a single-use assay reagent and provides sufficient volume to dispense 48 assay wells of HCT-Pgp cells into one 96-well plate. If desired, the two aliquots of HCT-Pgp cells may be thawed, combined to generate a single cell suspension, and then dispensed into all wells of an assay plate. The kit contains two aliquots of HCT-Pgp cells and two black, sterile, collagen-coated 96-well assay plates. In addition to HCT-Pgp cells and assay plates, this kit includes Cell Recovery Medium (CRM-p) used to perform a rapid thaw of the -80°C HCT-Pgp cells, Compound Screening Medium supplemented with daunorubicin (CSM+DR) used to prepare drug treatment media, Wash Buffer, two aliquots of Lysis Reagent, and the potent reference inhibitor Tariquidar. HCT-Pgp cells are a proprietary cell line derived from human colorectal adenocarcinoma cells that have been extensively selected for high-level expression of native P-glycoprotein. The primary application of this MDR1 drug interaction assay kit is to rapidly assess drug candidates as either inhibitors, substrates, or non-substrates of P-glycoprotein. This is accomplished by co-treating the cells with varying concentrations of the test drug and a fixed concentration of daunorubicin, a well-characterized fluorescent substrate for P-glycoprotein mediated transport. Protein function: Energy-dependent efflux pump responsible for decreased drug accumulation in multidrug-resistant cells. [The UniProt Consortium]