KV1.3 -Port-a-Patch - "Automated internal perfusion of Jurkat cells expressing KV1.3 channels"
Port-a-Patch application note: (0.2 MB)
KV1.3 is a voltage-gated potassium channel which plays a role in human T cell activation and proliferation, cell-mediated cytotoxicity and volume regulation. It is therefore an important target for the therapeutic control of T cell
responses. The KV1.3 channel is endogenously expressed in Jurkat cells, an immortalised T lymphocyte cell line. The channel is activated by membrane depolarisation at voltages typically more positive than -40 mV with rapid activation kinetics and slower inactivation. In common with many other voltage-gated potassium channels, KV1.3 can be blocked by replacing K+ with Cs+ ions in the pipette solution. Using Nanion’s Internal Perfusion System, we were able to record stable KV1.3 currents from Jurkat cells and reliably block this potassium current by perfusing the internal side of the chip with a Cs+-containing solution. The KV1.3 mediated current could also be completely recovered by perfusing the inside of the chip with K+-containing solution. KV1.3 mediated currents can also be blocked by internal perfusion with tetraethylammonium (TEA). We present data here showing block of KV1.3 by increasing concentrations
of TEA using the Internal Perfusion System. The IC50 that we obtained for TEA applied internally was in good agreement with the literature.