Chapter title |
Methods for Using a Genetically Encoded Fluorescent Biosensor to Monitor Nuclear NAD +
|
---|---|
Chapter number | 26 |
Book title |
ADP-ribosylation and NAD+ Utilizing Enzymes
|
Published in |
Methods in molecular biology, August 2018
|
DOI | 10.1007/978-1-4939-8588-3_26 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8587-6, 978-1-4939-8588-3
|
Authors |
Michael S. Cohen, Melissa L. Stewart, Richard H. Goodman, Xiaolu A. Cambronne, Cohen, Michael S., Stewart, Melissa L., Goodman, Richard H., Cambronne, Xiaolu A. |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 4 | 24% |
Student > Bachelor | 3 | 18% |
Professor | 1 | 6% |
Professor > Associate Professor | 1 | 6% |
Other | 0 | 0% |
Unknown | 4 | 24% |
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Immunology and Microbiology | 1 | 6% |
Other | 0 | 0% |
Unknown | 5 | 29% |