Protocol

High-Throughput Measurement of the Ca2+-Dependent ATPase Activity in COS Microsomes

  1. Peter Vangheluwe1
  1. Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, B3000 Leuven, Belgium

    Abstract

    We provide a detailed procedure to determine the Ca2+-dependent ATPase activity in COS or HEK293 cells overexpressing a Ca2+ 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.

    Footnotes

    • ↵1 Correspondence: peter.vangheluwe{at}med.kuleuven.be

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    1. doi:10.1101/pdb.prot076885 Cold Spring Harb Protoc 2014: pdb.prot076885- © 2014 Cold Spring Harbor Laboratory Press
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