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A number of recent studies have described the activation of BK(Ca) channels by steroid hormones such as estrogen. The proposed mechanisms are diverse and include both the direct interaction with the ion channel subunits and the stimulation via receptor activation and cell signalling pathways. To investigate the activation of BK(Ca) channels by estrogen we devised a cell-free system by incorporating recombinant channels of known subunit composition into artificial bilayers and recorded single channel currents. This chapter describes the methods used to prepare purified membrane fractions from cultured cells and the construction of artificial phospholipids bilayers for the incorporation and recording of ion channels.

Original publication

DOI

10.1007/978-1-59745-526-8_14

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2008

Volume

491

Pages

177 - 186

Keywords

Cell Line, Electrophysiology, Estrogens, Humans, Kidney, Large-Conductance Calcium-Activated Potassium Channel alpha Subunits, Lipid Bilayers, Phosphatidylethanolamines, Phosphatidylserines, Signal Transduction