For Research Use Only
NAD+ is supplied exclusively for in vitro laboratory research. Not for human or veterinary use. No dosing, administration, or reconstitution guidance is provided.
NAD+ 1000mg — Nicotinamide Adenine Dinucleotide for Metabolic, Sirtuin, and DNA Repair Research
NAD+ 1000mg is a research-grade preparation of nicotinamide adenine dinucleotide (NAD+), the oxidised form of the essential coenzyme that participates in over 500 enzymatic reactions in human metabolism. NAD+ serves as a hydride acceptor in catabolic reactions (glycolysis, the TCA cycle, and beta-oxidation), a substrate for NAD+-consuming enzymes including sirtuins (SIRT1–7), poly(ADP-ribose) polymerases (PARPs), and CD38/CD157 ectoenzymes, and a precursor for cyclic ADP-ribose (cADPR) — a second messenger involved in calcium signalling. Each vial supplied by AbsoluteBioLab contains 1000mg of NAD+ (free acid form), verified by HPLC to ≥99.0% purity with identity confirmed by UV spectroscopy (λmax 260 nm) and LC-MS.[1]
NAD+ is a dinucleotide coenzyme, not a peptide. It is distinct from NADH (the reduced form), NADP+ (the phosphorylated form), NMN (nicotinamide mononucleotide, a biosynthetic precursor), and NR (nicotinamide riboside, a salvage pathway precursor). AbsoluteBioLab supplies NAD+ as the free acid form, suitable for in vitro cell culture, biochemical assay, and enzyme kinetics research applications.
Compound Identity
| Attribute | Value | Notes |
|---|---|---|
| Full Name | Nicotinamide Adenine Dinucleotide (oxidised) | Free acid form |
| Abbreviation | NAD+, NAD, DPN | Diphosphopyridine nucleotide (historical) |
| CAS Number | 53-84-9 | NAD+ free acid |
| Molecular Formula | C21H27N7O14P2 | Free acid |
| Molecular Weight | 663.4 Da | Free acid |
| UV Absorbance | λmax 260 nm (ε = 18,000 M⁻¹cm⁻¹) | Adenine chromophore; used for quantification |
| Primary Functions | Redox coenzyme, sirtuin substrate, PARP substrate | 500+ enzymatic reactions |
| Key Consuming Enzymes | SIRT1–7, PARP1/2, CD38, NAMPT | NAD+ is cleaved, not reduced, by these enzymes |
| Physical Form | White to off-white lyophilised powder | Free acid; hygroscopic |
Analytical Specification and Release Testing
| Test Parameter | Method | Specification |
|---|---|---|
| Purity (HPLC) | C18 column, UV 260 nm, area normalisation | ≥99.0% |
| Identity (LC-MS) | LC-MS/MS | m/z 664.1 [M+H]+ — conforms |
| UV Absorbance | UV spectroscopy | λmax 260 nm — conforms |
| Residual Moisture | Karl Fischer titration | <5.0% w/w |
| Appearance | Visual inspection | White to off-white powder — conforms |
Analytical Note — AbsoluteBioLab Quality Team
NAD+ is characterised by HPLC with UV detection at 260 nm, exploiting the adenine chromophore (ε = 18,000 M⁻¹cm⁻¹). This wavelength is specific to the adenine ring system and distinguishes NAD+ from nicotinamide mononucleotide (NMN) and nicotinamide riboside (NR), which lack the adenine chromophore at this wavelength. LC-MS identity confirmation uses the [M+H]⁺ ion at m/z 664.1, consistent with the free acid molecular weight of 663.4 Da. Batch-specific CoA documents are available through the CoA Portal.
Biology and Research Context — NAD+ in Metabolism, Sirtuin Signalling, and DNA Repair
NAD+ occupies a unique position in cellular biology as both a redox coenzyme and a signalling molecule substrate. As a redox coenzyme, NAD+ accepts hydride equivalents (H⁻) from metabolic substrates during glycolysis (glucose → pyruvate), the TCA cycle (isocitrate → α-ketoglutarate, malate → oxaloacetate), and beta-oxidation (acyl-CoA → enoyl-CoA), generating NADH. NADH is then reoxidised to NAD+ by the mitochondrial electron transport chain (Complex I), coupling NADH oxidation to ATP synthesis via oxidative phosphorylation. The NAD+/NADH ratio is therefore a direct readout of the cell’s metabolic and redox state, and perturbations in this ratio are associated with metabolic dysfunction, mitochondrial disease, and cellular stress responses.[1]
As a substrate for NAD+-consuming enzymes, NAD+ is cleaved (not reduced) by sirtuins (SIRT1–7), poly(ADP-ribose) polymerases (PARP1/2), and CD38/CD157 ectoenzymes. Sirtuins are NAD+-dependent protein deacylases that regulate metabolic gene expression, mitochondrial biogenesis, DNA repair, and cellular stress responses. PARP1 and PARP2 consume NAD+ during DNA damage repair, and excessive PARP activation under conditions of genotoxic stress can deplete cellular NAD+ pools, impairing sirtuin function and mitochondrial metabolism. CD38 is the primary NAD+-consuming enzyme in mammals and its expression increases with inflammatory stimuli, contributing to changes in NAD+ pool dynamics.[2]
Intracellular NAD+ levels are regulated by the balance between biosynthesis (via the salvage pathway, de novo synthesis from tryptophan, and the Preiss-Handler pathway) and consumption by NAD+-dependent enzymes. The rate-limiting enzyme in the salvage pathway is NAMPT (nicotinamide phosphoribosyltransferase), which converts nicotinamide to NMN. Research into NAD+ biology has characterised the role of NAMPT activity, CD38 expression, and PARP activation in regulating cellular NAD+ pools under conditions of metabolic stress, genotoxic challenge, and inflammatory stimulation in pre-clinical models.[3]
Research Applications
Sirtuin Biochemistry and Enzyme Kinetics
NAD+ is the essential co-substrate for all seven mammalian sirtuins (SIRT1–7). In vitro sirtuin activity assays — including fluorometric SIRT1 activity assays, SIRT3 deacetylase assays, and SIRT6 deacylase assays — require NAD+ as a co-substrate at defined concentrations. AbsoluteBioLab’s NAD+ 1000mg is suitable for use in sirtuin biochemistry research, enzyme kinetics studies, and high-throughput screening assays for sirtuin modulators.[2]
PARP Biology and DNA Damage Research
PARP1 and PARP2 consume NAD+ to synthesise poly(ADP-ribose) chains on target proteins during DNA damage repair. NAD+ is used as a substrate in PARP activity assays, NAD+ depletion studies, and research into the relationship between DNA damage, PARP activation, and NAD+ metabolism. These applications are relevant to cancer biology, genotoxicity research, and the development of PARP inhibitors.[3]
Metabolic Research and NAD+/NADH Ratio Studies
NAD+ is used in cell-free metabolic assays to study glycolytic enzyme kinetics (lactate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase), TCA cycle enzyme activity (malate dehydrogenase, isocitrate dehydrogenase), and beta-oxidation. NAD+/NADH ratio measurement is a standard readout of cellular metabolic state, and NAD+ is used as a reference standard in NAD+/NADH quantification assays.
Cellular Senescence and Metabolic Research
NAD+ is used as a tool compound in cellular senescence research to investigate the relationship between NAD+ pool dynamics, sirtuin activity, mitochondrial function, and metabolic gene expression in cell culture models of replicative senescence and oxidative stress-induced cellular stress. In pre-clinical cell-based models, NAD+ supplementation has been shown to modulate sirtuin activity and mitochondrial function markers — findings that support its use as a research tool in metabolic biology programmes.[2] [3]
Storage and Transport
| Condition | Specification |
|---|---|
| Long-Term Storage (lyophilised, sealed) | −20°C, desiccated, protected from light |
| Stability Period | Stated on the batch-specific CoA |
| Transit | Ambient; cold-chain packaging included |
Note: Aqueous solution stability, solubility, pH sensitivity, and reconstitution guidance are not provided, consistent with AbsoluteBioLab’s Research Use Only supply policy. NAD+ is hygroscopic and susceptible to hydrolysis in solution; researchers should refer to their institutional SOPs for handling dissolved preparations. Stability period is stated on the batch-specific CoA.
Batch Documentation
Every batch of NAD+ 1000mg dispatched by AbsoluteBioLab is accompanied by a batch-specific Certificate of Analysis confirming HPLC purity (≥99.0%), LC-MS identity, UV absorbance, and Karl Fischer moisture. CoA documents are available via the Certificate of Analysis Portal. For further information on our analytical methodology, refer to the guide on HPLC Purity in Research Compound Testing.
Frequently Asked Questions
What is the difference between NAD+, NADH, NMN, and NR?
NAD+ (nicotinamide adenine dinucleotide, oxidised form) and NADH (reduced form) are the two interconvertible forms of the NAD coenzyme. NAD+ accepts electrons during catabolism to become NADH, which is then reoxidised to NAD+ by the electron transport chain. NMN (nicotinamide mononucleotide) and NR (nicotinamide riboside) are NAD+ precursors — smaller molecules that are converted to NAD+ intracellularly via the NAD+ salvage pathway. For in vitro biochemical research requiring direct NAD+ as a substrate (e.g., sirtuin assays, PARP assays, enzyme kinetics), NAD+ free acid is the appropriate form. For cell culture studies investigating NAD+ metabolism and precursor uptake, NMN or NR may be more appropriate as they are cell-permeable NAD+ precursors.
Is a Certificate of Analysis available?
Yes. Batch-specific CoA documents — including annotated HPLC chromatograms and LC-MS data — are available through the AbsoluteBioLab CoA Portal and are included with each order.
What is the chemical form supplied?
AbsoluteBioLab supplies NAD+ as the free acid form (CAS 53-84-9), not the sodium salt (CAS 606-68-8) or disodium salt. The free acid form is appropriate for most in vitro biochemical applications. The molecular formula is C₂₁H₂₇N₇O₁₄P₂ and the molecular weight is 663.4 Da.
References
- Guarente L. Calorie restriction and sirtuins revisited. Genes Dev. 2013;27(19):2072–2085. doi:10.1101/gad.227439.113
- Imai S, Guarente L. NAD+ and sirtuins in aging and disease. Trends Cell Biol. 2014;24(8):464–471. doi:10.1016/j.tcb.2014.04.002
- Cantó C, Menzies KJ, Auwerx J. NAD+ metabolism and its roles in cellular processes during ageing. Cell Metab. 2015;22(1):31–53. doi:10.1016/j.cmet.2015.06.013
- Verdin E. NAD+ in aging, metabolism, and neurodegeneration. Science. 2015;350(6265):1208–1213. doi:10.1126/science.aac4854
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