Calcium uptake in crude tissue preparation

Philip A. Bidwell, Evangelia G. Kranias

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)161-170
Number of pages10
JournalMethods in Molecular Biology
Volume1377
DOIs
StatePublished - 2016
Externally publishedYes

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media New York 2016.

Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.

Keywords

  • Ca
  • Calcium affinity
  • Calcium uptake
  • K
  • Oxalate
  • Phospholamban
  • SERCA
  • Sarcolipin
  • V

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