| Assay Method Information | |
| | Patch clamp assay |
| Description: | HEK293 cells were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (D-MEM/F-12) supplemented with 10% fetal bovine serum, 100 U/mL penicillin G sodium, 100 ug/mL streptomycin sulfate and 500 ug/mL G418. Before testing, cells in culture dishes were washed twice with Hank's Balanced Salt Solution, treated with Accutase and re-suspended in the culture media ( 20 106 cells in 20 mL). Cells in suspension were allowed to recover for 10 minutes in tissue culture incubator set at 37 C. in a humidified 95% air: 5% CO2 atmosphere. Immediately before use in the SyncroPatch 384PE system (SP384PE), the cells were washed twice in extracellular buffer (HB-PS) to remove the culture medium and re-suspended in 20 mL of HB-PS. Extracellular buffer was loaded into the wells of Nanion 384-well Patch Clamp (NPC-384, 4 M) chip (60 uL per well). Then, cell suspension was pipetted into the wells (20 uL per well) of the chip. After establishment of the whole-cell configuration, membrane currents were recorded using patch clamp amplifier in the SP384PE system. |
| Affinity data for this assay | |
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