Advantages of formate dehydrogenase reaction for efficient NAD+ quantification in biological samples.
Analytical biochemistry
confidence
Key findings
Describes a fluorometric assay using formate dehydrogenase for NAD+ quantification in tissue extracts; no clinical or biological endpoints reported.
View source on PubMed (PMID 32562604) ↗
- Sample size
- Not reported
- Population
- Rat brain cortex and mitochondria tissue extracts
- Dosing
- Not applicable
- Duration
- Not reported
- Route
- Not applicable
- Blinding
- not_reported
- Controls
- none
- Drug class
- coenzyme
Full abstract
The medical significance of NAD+-dependent metabolic regulation acquires increasing attention, demanding rapid and clinically feasible quantification of NAD+ in complex biological samples. Here we describe the usage of formate dehydrogenase for a straightforward and highly specific fluorometric assay of NAD+ in tissue extracts, not requiring chromatographic separation of nucleotides. The assay employs the irreversible reaction of formate oxidation coupled to NAD+ reduction, catalyzed by the enzyme which has high affinity and specificity to NAD+, and is stable under a variety of conditions. The assay reliably quantifies NAD+ in the methanol extracts of the rat brain cortex and mitochondria.