Measuring the rate constants of Ca²⁺ indicators

Fluorescent Ca(2+) indicators are widely used to measure the concentration of free Ca(2+) ([Ca(2+)](free)) in biological processes. To determine the exact kinetics of changes in [Ca(2+)](free) and the processes underlying these changes (e.g., Ca(2+) binding to Ca(2+)-binding proteins), it is necessa...

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Bibliographic Details
Published inCold Spring Harbor protocols Vol. 2014; no. 7; p. 763
Main Authors Faas, Guido C, Mody, Istvan
Format Journal Article
LanguageEnglish
Published United States 01.07.2014
Subjects
Calcium - analysis
Calcium - chemistry
Chemistry Techniques, Analytical
Fluorescent Dyes - chemistry
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Summary:Fluorescent Ca(2+) indicators are widely used to measure the concentration of free Ca(2+) ([Ca(2+)](free)) in biological processes. To determine the exact kinetics of changes in [Ca(2+)](free) and the processes underlying these changes (e.g., Ca(2+) binding to Ca(2+)-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 (k(on) and k(off)) of Ca(2+) indicators.
ISSN:1559-6095
DOI:10.1101/pdb.prot073288