Whole-cell patch-clamp recording of recombinant voltage-sensitive Ca2+ channels heterologously expressed in HEK-293
María A Gandini1, Alejandro Sandoval, Ricardo Felix
1Department of Cell Biology, Center for Research and Advanced Studies of the National Polytechnic Institute (Cinvestav-IPN), Mexico City, Mexico;
Abstract:
Several factors, including showing a high efficiency of transfection and protein production, faithful translation and processing of proteins, and possession of a small cell size, have together made human embryonic kidney 293 (HEK-293) cells a popular choice among electrophysiologists. This cell line has been extensively used to study recombinant heterooligomeric voltage-sensitive Ca(2+) channels. The main reasons for this are that the properties of the channel complexes can be studied in isolation and that it is also possible to define the subunit composition of the expressed channels. This protocol provides a detailed method for the transfection of, and obtaining electrophysiological data from, HEK-293 cells transiently expressing neuronal (N-type) recombinant Ca(2+) channels.
