White clear bottomed poly d lysine coated 96 well plates

HEK-293T cells were maintained in DMEM/high glucose/L-glutamine/HEPES supplemented with 1 mM sodium pyruvate (Life Technologies) and 10% FBS (PAA Laboratories) at 37 °C in 5% CO₂. To express URAT1, plasmids were reverse transfected into HEK-293T cells. DNA (10 μg) was mixed with 30 μl of DreamFect Gold (Boca Scientific) in 2 ml OptiMem (Life Technologies). After 20 minutes, 18 ml of media containing 2 × 10⁷ cells in suspension was added and mixed, and 200 μl per well were plated onto white clear-bottomed poly-D-lysine coated 96-well plates (BD Biosciences). The cells were assayed the following day.

HEK-293T cells were maintained in DMEM/high glucose/L-glutamine/HEPES supplemented with 1 mM sodium pyruvate (Life Technologies) and 10% FBS (PAA Laboratories) at 37 °C in 5% CO₂. To express URAT1, OAT4 or OAT1, plasmids were reverse transfected into HEK-293T cells. DNA (10 µg) was mixed with 30 µl of DreamFect Gold (Boca Scientific) in 2 ml OptiMem (Life Technologies). After 20 minutes, 18 ml of media containing 2 × 10⁷ cells in suspension was added and mixed, and 200 µl per well were plated onto white clear-bottomed poly-D-lysine coated 96-well plates (BD Biosciences). The cells were assayed the following day.

HEK-293T cells were maintained in Dulbecco’s Modified Eagle’s Medium/high glucose/l-glutamine/2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) supplemented with 1 mM sodium pyruvate (Life Technologies) and 10% fetal bovine serum (GE Healthcare Life Sciences), at 37 °C in 5% CO₂. Plasmids were reverse transfected into HEK-293T cells. Ten micrograms of DNA was mixed with 30 µl of DreamFect Gold (Boca Scientific) in 2 ml OptiMem (Life Technologies). After 20 min, 18 ml of media containing 2E7 cells in suspension was added and mixed, and 200 µl per well was plated onto white clear-bottomed poly-d-lysine coated 96-well plates (BD Biosciences). The cells were assayed the next day.