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Calcium Uptake in Crude Tissue Preparation

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P-Type ATPases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1377))

Abstract

The various isoforms of the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) are responsible for the Ca2+ uptake from the cytosol into the endoplasmic or sarcoplasmic reticulum (ER/SR). In some tissues, the activity of SERCA can be modulated by binding partners, such as phospholamban and sarcolipin. The activity of SERCA can be characterized by its apparent affinity for Ca2+ as well as maximal enzymatic velocity. Both parameters can be effectively determined by the protocol described here. Specifically, we describe the measurement of the rate of oxalate-facilitated 45Ca uptake into the SR of crude mouse ventricular homogenates. This protocol can easily be adapted for different tissues and animal models as well as cultured cells.

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Acknowledgement

This work was supported by NIH grants HL-26057 and HL-64018 to E. G. K. and AHA grant 13POST13860006 to P. A. B.

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Authors and Affiliations

  1. Department of Pharmacology and Cell Biophysics, College of Medicine, University of Cincinnati, 231 Albert Sabin Way, Cincinnati, OH, 45267-0575, USA

    Philip A. Bidwell & Evangelia G. Kranias Ph.D.

Authors
  1. Philip A. Bidwell
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  2. Evangelia G. Kranias Ph.D.
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Corresponding author

Correspondence to Evangelia G. Kranias Ph.D. .

Editor information

Editors and Affiliations

  1. Department of Biochemistry, Biocenter, University of Oxford, Oxford, Oxfordshire, United Kingdom

    Maike Bublitz

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Bidwell, P.A., Kranias, E.G. (2016). Calcium Uptake in Crude Tissue Preparation. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_16

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  • DOI: https://doi.org/10.1007/978-1-4939-3179-8_16

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3178-1

  • Online ISBN: 978-1-4939-3179-8

  • eBook Packages: Springer Protocols

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