Protocol to study human Kv1.2 potassium channel pathogenic sequence variants using two-electrode voltage-clamp technique.

The abietane diterpenoid pisiferic acid (PA) from conifer Chamaecyparis pisifera is a pan-rescuer of human Kv1.2 channel pathogenic loss-of-function (LOF) sequence variants. Here, we present a protocol for the study of Kv1.2 channel sequence variants using the Xenopus laevis oocyte expression system and two-electrode voltage-clamp (TEVC) electrophysiology. We describe steps for studying mutant Kv1.2, wild-type Kv1.2, and wild-type Kv1.1 cRNA combinations in Xenopus laevis oocytes; performing TEVC; and data analysis. For complete details on the use and execution of this protocol, please refer to Manville et al.1.