title: High-throughput measurement of the Ca2+-dependent ATPase activity in COS microsomes.
abstract: We provide a detailed procedure to determine the Ca(2+)-dependent ATPase activity in COS or HEK293 cells overexpressing a Ca(2+) pump. The ATPase activity is determined by the Baginsky method, which allows measurement of the steady-state production of inorganic phosphate (Pi). We have adapted this widely applied method into a sensitive, fast, and semi-high-throughput protocol suitable for use in a 96-well plate format.
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Ilse
authors.initials: I
authors.lastname: Vandecaetsbeek
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Tine
authors.initials: T
authors.lastname: Holemans
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Frank
authors.initials: F
authors.lastname: Wuytack
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Vangheluwe
mesh_terms: D000818:Animals; D019556:COS Cells; D000252:Calcium-Transporting ATPases / Q000032:analysis*; D002522:Chlorocebus aethiops; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008861:Microsomes / Q000201:enzymology* / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D010710:Phosphates; D000252:Calcium-Transporting ATPases