Assay Report Card

ID: ALA3706160

Type: Binding

Description: Electrophysiology Assay: Block of Kir1.1 (ROMK1) currents was examined by whole cell voltage clamp (Hamill et. al. Pfluegers Archives 391:85-100 (1981)) using the IonWorks.RTM. Quattro automated electrophysiology platform (Molecular Devices, Sunnyvale, Calif.). Chinese hamster ovary cells stably expressing Kir1.1 channels were maintained in T-75 flasks in cell culture media in a humidified 10% CO.sub.2 incubator at 37.degree. C. Prior to an experiment, Kir1.1 expression was induced by overnight incubation with 1 mM sodium butyrate. On the day of the experiment, cells were dissociated with 2.5 ml of Versene.TM. (Invitrogen 15040-066) for approximately 6 minutes at 37.degree. C. and suspended in 10 ml of bath solution containing (in mM): 150 NaCl, 10 KCl, 2.7 CaCl.sub.2, 0.5 MgCl.sub.2, 5 HEPES, pH 7.4. After centrifugation, the cell pellet was resuspended in approximately 4.0 ml of bath solution and placed in the IonWorks.RTM. instrument. The intracellular solution consisted of (in mM): 80 K gluconate, 40 KCl.

Organism: Homo sapiens

Cell type: ovary