The MDR1/P-gp Ligand Screening Kit is designed for rapidly screening test compounds for modulation of efflux transporter activity in MDR1-expressing cell lines. The assay uses a lipophilic non-fluorescent P-gp substrate that readily diffuses through the plasma membrane, where it is hydrolyzed to an active fluorophore by cytosolic esterases. The resulting hydrophilic fluorophore is neither membrane permeable nor a substrate for P-gp, hence it remains trapped inside the cell. In MDR1-expressing cell lines, the lipophilic pro-fluorophore is continuously extruded from the cytosol by P-gp, leading to a low intracellular fluorescence. Inhibition of P-gp-mediated efflux by a test compound leads to increased intracellular fluorescence. Specific transporter activity is quantified by comparison with fluorescence accumulated in the presence and absence of a saturating concentration of the included selective P-gp inhibitor. The assay is highly sensitive, has a simple no-wash protocol and is highthroughput adaptable. The kit contains a complete set of reagents sufficient for performing 100 reactions in a 96-well plate format.
Size
100 assays
Concentration
n/a
Applications
Screening and characterization of drugs and new chemical entities for inhibition of native/recombinant MDR1/P-gp efflux transporter.Identification/characterization of cells and cell lines with a high level of MDR1-mediated efflux (i.e. chemotherapy-resistant).