Description
PrymaLab · Research Use Only
NAD+ 1000mg
Nicotinamide adenine dinucleotide · gram-scale vial · 663.43 Da
NAD+ 1000mg is a gram-scale lyophilized vial of nicotinamide adenine dinucleotide, CAS 53-84-9, molecular weight 663.43. One gram is approximately 1.51 millimoles, a quantity suited to enzyme assays and biochemical work where the coenzyme is a stoichiometric reagent rather than a trace additive.
Specification Table
| Property | Value |
|---|---|
| Compound | Nicotinamide adenine dinucleotide, oxidised form |
| Vial content | 1000 mg lyophilized powder |
| Molar content of vial | Approximately 1.51 mmol |
| Stock at 20 ml diluent | Approximately 75.4 mM |
| Common designation | NAD+ |
| CAS number | 53-84-9 |
| Molecular formula | C21H27N7O14P2 |
| Molecular weight | 663.43 g/mol |
| Compound class | Dinucleotide coenzyme. Not a peptide |
| Structure | Nicotinamide mononucleotide joined to adenosine monophosphate through a pyrophosphate bridge |
| Redox partner | NADH, the reduced form |
| Principal enzyme classes consuming NAD+ | Sirtuins, PARPs, CD38, SARM1 |
| Principal biosynthetic routes | De novo from tryptophan, Preiss-Handler from nicotinic acid, salvage from nicotinamide |
| Rate-limiting salvage enzyme | NAMPT, nicotinamide phosphoribosyltransferase |
| Appearance | White to off-white lyophilized powder |
| Purity | Per lot-specific certificate of analysis |
| Solubility | Freely water soluble |
| Aqueous stability | Degrades in aqueous solution, faster at alkaline pH and at raised temperature |
| Storage, lyophilized | -20°C, protected from light and moisture |
| Storage, reconstituted | 2-8°C, protected from light, short hold only |
| Regulatory status | No approved human or veterinary injectable formulation |
How Is NAD+ Made in Cells?
Three separate routes converge on the same molecule, and knowing which one a given experimental system relies on determines what a precursor addition will and will not do.
The de novo pathway starts from tryptophan and proceeds through the kynurenine branch to quinolinic acid and onward, and it is the only route that creates new pyridine ring, and it is comparatively slow and tissue-restricted, with liver contributing most.
The Preiss-Handler pathway starts from nicotinic acid, converting it through nicotinic acid mononucleotide and nicotinic acid adenine dinucleotide before amidation. It is shorter than de novo synthesis and depends on dietary nicotinic acid availability.
The salvage pathway starts from nicotinamide, the product released by every NAD+-consuming enzyme. NAMPT converts nicotinamide to nicotinamide mononucleotide, and NMNAT completes the conversion. This route carries most of the flux in most tissues.
NAMPT is rate-limiting in the salvage route, which is why it appears so often in the literature and why NAMPT inhibitors such as FK866 are the standard tool for depleting cellular NAD+ experimentally.
How Do Precursors Compare With the Coenzyme Itself?
A recurring question in this area is why anyone would supply a precursor when the coenzyme is available, and the answer is about membrane permeability rather than potency.
NAD+ itself crosses the plasma membrane poorly. It is large, carries multiple negative charges from its pyrophosphate, and has no widely characterised dedicated transporter in mammalian plasma membranes. Adding it to culture medium does not straightforwardly raise intracellular concentration.
Nicotinamide mononucleotide and nicotinamide riboside are both smaller and are proposed to enter cells more readily, NR through nucleoside transporters after which NRK phosphorylates it. Whether NMN enters intact or is first dephosphorylated to NR extracellularly has been actively debated in the literature.
Nicotinamide itself enters freely, being small and uncharged, but it also inhibits sirtuins at raised concentration, which complicates its use as a way of supporting sirtuin activity.
For enzyme assays in cell-free systems, none of this applies. The coenzyme is added directly to the reaction and permeability is irrelevant, which is where a gram-scale vial finds most of its use.
What Analytical Verification Suits a Gram-Scale Purchase?
NAD+ 1000mg justifies more verification than a smaller fill, and NAD+ offers unusually convenient analytical handles.
Ultraviolet absorbance gives a direct concentration readout. The oxidised form absorbs strongly at 260 nm with an extinction coefficient around 18,000 per molar per centimetre. Its reduced counterpart shows a second peak near 340 nm that the oxidised form lacks, so the 260 to 340 relationship reports on the redox state of the preparation.
An enzymatic cycling assay offers the most sensitive route where quantities are small, coupling NAD+ to a colorimetric or fluorescent readout through alcohol dehydrogenase or a similar system. It also distinguishes functional coenzyme from degraded material that still absorbs.
High-performance liquid chromatography resolves NAD+ from its degradation products and from nicotinamide, which matters for a preparation that has been stored or shipped under uncertain conditions.
Water content deserves attention at gram scale. A hygroscopic lyophilized powder that has taken up moisture weighs more than the coenzyme it contains, and at gram scale the arithmetic error becomes material.
How Should a Gram of NAD+ 1000mg Be Handled?
With NAD+ 1000mg the case is stronger still for dividing the dry powder rather than making one large solution.
Dry material at -20°C in a sealed container protected from moisture is stable for extended periods. The same material in aqueous solution is not. Every gram reconstituted at once starts a clock on the entire purchase.
Weigh out portions matching planned consumption and return the bulk to cold storage promptly. Minimise the time the container spends open, since the powder is hygroscopic and ambient humidity is the enemy of both weight accuracy and long-term stability.
Allow the sealed container to reach room temperature before opening. Opening a cold container in a humid room condenses water directly onto the powder, which is the single most common way a well-stored gram-scale reagent is quietly ruined.
Where solution is needed, prepare in lightly acidic buffer, aliquot into single-use volumes, freeze immediately, and thaw each aliquot once.
How Does Compartment Access Constrain Interpretation?
Adding a coenzyme to a system is not the same as raising its concentration where the question lies, and this distinction defeats more NAD+ experiments than any other.
In a cell-free enzyme assay there is one compartment and the added concentration is the concentration acting, and that simplicity is exactly why gram-scale material finds most of its use in biochemistry rather than cell biology.
In a cell-culture system there are at least three relevant compartments: the medium, the cytosol with the nucleus, and the mitochondrial matrix. Added coenzyme reaches the first freely, the second poorly, and the third not at all through direct transit.
What may happen instead is extracellular degradation. Ectoenzymes including CD38 and various nucleotidases act on added coenzyme at the cell surface, and the products, including nicotinamide riboside and nicotinamide, do enter cells.
That means an apparent intracellular effect of added coenzyme may in fact be an effect of its extracellular breakdown products. Distinguishing the two requires either measuring the extracellular species over time or comparing against direct addition of the candidate products.
A study reporting that added coenzyme raised intracellular concentration, without addressing the degradation route, has not excluded the simpler explanation.
What Does the Salvage Route Mean for Depletion Experiments?
Experiments that reduce cellular coenzyme concentration are common, and the salvage pathway determines how well they work.
NAMPT inhibition using FK866 is the standard approach, and it works because salvage carries most of the flux in most cell types. Blocking the rate-limiting step causes concentration to fall over hours as consumption continues without replacement.
The kinetics are informative in themselves. How fast concentration falls after NAMPT inhibition reports on the consumption rate in that cell type, since the decline reflects turnover once resupply is blocked.
The approach fails where an alternative route is active. Cells with functional Preiss-Handler capacity can use nicotinic acid to bypass the block entirely, so medium composition determines whether the inhibitor achieves depletion at all.
That is a frequent and avoidable confound. Culture medium formulations differ in their pyridine content, and a medium supplying nicotinic acid provides an escape route that a nicotinamide-only medium does not.
Stating the medium formulation, and ideally testing depletion directly rather than assuming the inhibitor worked, is what makes a depletion experiment interpretable.
What Scale of Work Does NAD+ 1000mg Match?
A gram is a lot of coenzyme. Matching it to the right kind of work avoids buying material that degrades before it is used.
Enzyme kinetics is the clearest fit. Determining a Michaelis constant requires a concentration series spanning at least an order of magnitude either side of the expected value, run in replicate, and each curve consumes substrate in quantity.
Coupled assay systems consume more still, since the coenzyme is both the analyte and the reporter in many of them, and NAD+ 1000mg supports the number of wells that a proper screen requires.
Structural work on coenzyme-binding enzymes is the other fit, since co-crystallisation and soaking experiments both use material at concentrations far above assay levels.
Cell-culture programmes rarely justify a gram. The permeability problem means most of what is added never reaches the compartment of interest, so buying at this scale for culture work purchases degradation rather than data.
Reconstitution and Storage in Laboratory Practice
Twenty millilitres of diluent gives NAD+ 1000mg at approximately 75.4 mM, matching the concentration a 500mg vial in 10 ml produces, which keeps dilution arithmetic transferable between the two formats.
Dissolution is rapid. The compound is freely water soluble and needs no coaxing, so any resistance to dissolving is worth investigating rather than overcoming.
Keep solutions cold and dark from preparation onward, and treat elapsed time in solution as a variable to record rather than ignore.
Hold dry material at -20°C, sealed, dark, and dry. Record lot, water content where determined, the mass weighed, diluent identity and pH, the concentration obtained measured rather than assumed, and the date. At gram scale spread across many experiments, that record is what allows a discrepant finding to be traced.
Published Literature
References verified against the publisher record. NAD+ biology rests on a substantially larger and more independent evidence base than most compounds in this catalogue.
- Cantó C, Menzies KJ, Auwerx J. Cell Metabolism. 2015;22(1):31-53.
- Verdin E. Science. 2015;350(6265):1208-1213.
- Yoshino J, Baur JA, Imai SI. Cell Metabolism. 2018;27(3):513-528.
- Rajman L, Chwalek K, Sinclair DA. Cell Metabolism. 2018;27(3):529-547.
- Cambronne XA, Kraus WL. Trends in Biochemical Sciences. 2020;45(10):858-873.
- Bogan KL, Brenner C. Annual Review of Nutrition. 2008;28:115-130.
Frequently Asked Questions
What is NAD+ 1000mg?
A gram-scale lyophilized vial of nicotinamide adenine dinucleotide, CAS 53-84-9, molecular weight 663.43. One gram is approximately 1.51 millimoles, suited to enzyme assays where the coenzyme is a stoichiometric reagent rather than a trace additive.
What are the three biosynthetic routes?
De novo from tryptophan through the kynurenine branch, the only path creating new pyridine ring. Preiss-Handler from nicotinic acid, which is shorter and diet-dependent. And salvage from nicotinamide, which carries most of the flux in most tissues.
Why does NAMPT appear so often?
Because it is rate-limiting in the salvage pathway, which handles most NAD+ turnover. That also makes it the standard experimental handle, and NAMPT inhibitors such as FK866 are the conventional tool for depleting cellular NAD+ in a controlled way.
Why supply precursors rather than the coenzyme?
Membrane permeability, not potency. NAD+ crosses the plasma membrane poorly, being large and multiply charged with no widely characterised mammalian plasma membrane transporter. Adding it to culture medium does not straightforwardly raise intracellular concentration.
How do NMN and NR differ?
Both are smaller than the coenzyme and proposed to enter cells more readily. NR enters through nucleoside transporters and is then phosphorylated by NRK, and whether NMN enters intact or is dephosphorylated to NR extracellularly first has been actively debated.
Why is nicotinamide itself complicated?
It enters cells freely, being small and uncharged, but it also inhibits sirtuins at raised concentration. That makes it awkward as a way of supporting sirtuin activity, since the same addition that supplies substrate also suppresses the enzyme.
Does permeability matter in cell-free assays?
No. In a cell-free enzyme assay the coenzyme is added directly to the reaction mixture and permeability is irrelevant, which is where a gram-scale vial finds most of its practical use rather than in cell-culture work.
How is the redox state checked?
Through the relationship between two absorbance peaks. The oxidised form absorbs strongly at 260 nanometres, while the reduced form shows an additional peak near 340 nanometres that the oxidised form lacks, so the two together report on preparation state.
Why does water content matter at gram scale?
Because a hygroscopic lyophilized powder that has taken up moisture weighs more than the coenzyme it contains. At milligram scale the error is tolerable. At gram scale, across many experiments drawing from one weighing, it becomes material.
What is the most common storage mistake?
Opening a cold sealed container in a humid room, which condenses water directly onto hygroscopic powder. Allowing the container to reach room temperature before opening avoids it, and it is the single easiest way to ruin a well-stored gram-scale reagent.
Compliance Statement
NAD+ 1000mg is sold exclusively for laboratory research use. It is not a drug, food, or cosmetic product, and it is not a dietary product of any kind. It is not approved by the FDA or any comparable authority for human or veterinary use, and no approved injectable formulation of this compound exists. This product is not intended to diagnose, treat, cure, or prevent any disease. It must not be given to humans or animals. Purchase is restricted to qualified researchers and institutions operating within applicable laws. All handling is the responsibility of the purchasing laboratory.
Other formats of NAD
NAD is also stocked as NAD+ Nasal Spray, NAD+ (injectable) 500mg/ml preloaded 3ml pen, NAD+ 100mg/ml preloaded 3ml pen and NAD+ 500mg. Each listing states its own quantity and concentration, and the pen and vial comparison explains what changes between formats.
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