Protocol

Measuring the Rate Constants of Ca2+ Indicators

  1. Istvan Mody1,2
  1. 1Department of Neurology, David Geffen School of Medicine, UCLA, Los Angeles, California 90095;
  2. 2Department of Physiology, David Geffen School of Medicine, UCLA, Los Angeles, California 90095

    Abstract

    Fluorescent Ca2+ indicators are widely used to measure the concentration of free Ca2+ ([Ca2+]free) in biological processes. To determine the exact kinetics of changes in [Ca2+]free and the processes underlying these changes (e.g., Ca2+ binding to Ca2+-binding proteins), it is necessary to know the exact binding properties of the indicator used. Here, we describe how to determine the binding rate constants (kon and koff) of Ca2+ indicators.

    Footnotes

    • ↵3 Correspondence: gfaas{at}ucla.edu

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    1. doi:10.1101/pdb.prot073288 Cold Spring Harb Protoc 2014: pdb.prot073288- © 2014 Cold Spring Harbor Laboratory Press
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    1. Fluorescence
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