Abstract

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.

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