| Assay Method Information | |
| | Two-electrode Voltage Clamp Electrophysiology |
| Description: | Two-electrode voltage clamp (TEVC) recordings were performed on Xenopus oocytes at room temperature 3-6 days postinjection using an OC-725C TEVC amplifier (Warner Instruments, Hamden, CT). Glass electrodes had a tip resistance of 0.5-2.5 megaohms and were pulled from thin walled glass capillary tubes (World Precision Instruments, Hertfordshire, UK) using a PC-10 puller (Narishige, East Meadow, NY). Voltage and current electrodes were filled with 0.3 and 3 M KCl, respectively. During recordings, oocytes were placed in a recording chamber and perfused with the extracellular recording solution comprised of 90 mM NaCl, 1 mM KCl, 10 mM HEPES, 0.5 mM BaCl2, and 0.01 mM EDTA (pH 7.4 with NaOH). Current responses were recorded at a holding potential of -40 and -60 mV for GluN1/N2 and GluN1/N3 receptors, respectively. Compounds were dissolved in extracellular recording solution and applied to the oocyte by gravity-driven perfusion using a computer-controlled 8-modular valve positioner (Digital MVP, Hamilton, Reno, NV). |
| Affinity data for this assay | |
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