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Updated: Feb 6, 2026

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
Published on: February 4, 2016
Methods for Using a Genetically Encoded Fluorescent Biosensor to Monitor Nuclear NAD
Michael S Cohen1, Melissa L Stewart2, Richard H Goodman2
1Department of Physiology and Pharmacology, Program in Chemical Biology, Oregon Health and Science University, Portland, OR, USA.
Abstract:
Free nicotinamide adenine dinucleotide (NAD+) serves as substrate for NAD+-consuming enzymes. As such, the local concentration of free NAD+ can influence enzymatic activities. Here we describe methods for using a fluorescent, genetically-encoded sensor to measure subcellular NAD+ concentrations. We also include a discussion of the limitations and potential applications for the current sensor. Presented in this chapter are (1) guidelines for calibrating instrumentation and experimental setups using a bead-based method, (2) instructions for incorporating required controls and properly performing ratiometric measurements in cells, and (3) descriptions of how to evaluate relative and quantitative fluctuations using appropriate statistical methods for ratio-of-ratio measurements.
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