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High-throughput measurement of the Ca2+-dependent ATPase activity in COS microsomes
Ilse Vandecaetsbeek1, Tine Holemans1, Frank Wuytack1
1Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium.
Cold Spring Harbor Protocols
|August 3, 2014
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.
