title: Measurement of intracellular Ca2+ release in permeabilized cells using 45Ca2+.
abstract: This protocol describes a technique to measure Ca(2+) release from the nonmitochondrial intracellular Ca(2+) stores in monolayers of saponin-permeabilized cells cultured in 12-well 4-cm(2) clusters. The (45)Ca(2+)-flux technique described here can only be applied to cell types that still adhere to the plastic after exposing them to saponin. We describe the permeabilization procedure, the loading of the nonmitochondrial Ca(2+) stores with (45)Ca(2+), and the subsequent (45)Ca(2+) efflux.
authors.affiliation: Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, KU Leuven Campus Gasthuisberg O&N I, 3000 Leuven, Belgium.
authors.forename: Tomas
authors.initials: T
authors.lastname: Luyten
authors.forename: Geert
authors.initials: G
authors.lastname: Bultynck
authors.forename: Jan B
authors.initials: JB
authors.lastname: Parys
authors.forename: Humbert
authors.initials: H
authors.lastname: De Smedt
authors.forename: Ludwig
authors.initials: L
authors.lastname: Missiaen
mesh_terms: D002118:Calcium / Q000378:metabolism*; D002132:Calcium Radioisotopes / Q000378:metabolism*; D002460:Cell Line; D002463:Cell Membrane Permeability; D003470:Culture Media; D017136:Ion Transport
chemical_list: D002132:Calcium Radioisotopes; D003470:Culture Media; D002118:Calcium