NAD+ (500 mg vial)

Protocol Overview

Concise summary of the regimen.

Dosing & Reconstitution

IM is the preferred at home route during active rebuilds because muscle tissue tolerates the acidic load better than shallow subcutaneous tissue. Slow injection, roughly 30 to 60 seconds per mL, reduces sting, and any dose above 150 mg (90 units) is split across two injection sites. Subcutaneous administration is acceptable when IM is not feasible, but it burns more because NAD+ is inherently acidic at a pH near 3.5 to 4.0; pre-buffered product is preferred, larger doses are split across multiple smaller injections rather than concentrating volume, and bicarbonate is never mixed into the syringe because that changes solution chemistry with no pharmacokinetic dataset behind it. Sites rotate systematically: love handles at the lateral hip sting least, then outer thigh, while the abdomen is more vascular and stings more.

Results Timeline

Sequence described in the research record. Individual response varies.

• Week 1: subtle energy improvement is reported during the first week of loading.
• Weeks 1 to 4: sleep and cognitive changes appear across this window.
• Weeks 4 to 8: benefits become sustained through the active rebuild.
• Weeks 9 and beyond: dosing steps down to weekly IM, oral maintenance, or both.

Reconstitution Steps

1. Allow the lyophilized vial to reach room temperature before opening to minimize condensation.
2. Draw 3.0 mL bacteriostatic water with a sterile syringe; sodium chloride containing bacteriostatic water reduces welting.
3. Inject slowly down the vial wall; do not aim at the powder. Avoid foaming.
4. Gently swirl until fully dissolved (do not shake). Solution should be clear and colorless.
5. Label with the reconstitution date and concentration; refrigerate at 2 to 8 °C, protected from light. Use within 14 days and inspect before each use; discard if discoloration or precipitate appears.
6. At 166.67 mg/mL, 1 unit = 0.01 mL » 1.67 mg on a U-100 insulin syringe. 50 mg = 30 units; 100 mg = 60 units; 150 mg = 90 units; 250 mg = 150 units, split across two sites.

Storage Instructions

Proper storage preserves peptide quality.

• Lyophilized: store at minus 20 °C or below, or minus 80 °C for storage beyond a year, in a dry, dark environment. NAD+ powder is hygroscopic.
• Reconstituted: refrigerate at 2 to 8 °C and use within about 14 days. Protect from light, since UV degrades NAD+.

• Avoid repeated freezing and thawing of reconstituted solution.
• Let refrigerated vials warm to room temperature for about 10 to 15 minutes before injecting, since cold solution stings more, and inspect before each use: fresh NAD+ solution is clear and colorless.

Supplies Needed

The figures below are based on 8 to 12 weeks of active IM rebuild at 100 mg three times weekly, which is 300 mg per week, followed by weekly maintenance.

Peptide Vials (NAD+, 500 mg vial):

• Eight weeks of active dosing totals 2,400 mg, about 5 vials of 500 mg; 12 weeks totals 3,600 mg, about 8 vials; a further 12 weeks of maintenance at 100 mg weekly totals 1,200 mg, about 3 vials.

Insulin Syringes (U-100):

• Per week: 3 U-100 insulin syringes during the active rebuild, 1 per week during maintenance.
• Full cycle: 24 syringes for 8 weeks active (24 injections), 36 syringes for 12 weeks active (36 injections).

Bacteriostatic Water (10 mL bottles):

• Each 500 mg vial takes 3.0 mL, so one 10 mL bottle covers 3 vials: 2 bottles for 8 weeks active (5 vials) and 3 bottles for 12 weeks active (8 vials), with sodium chloride containing bacteriostatic water preferred to reduce welting.

Alcohol Swabs:

• Per injection: 2 swabs, one for the vial stopper and one for the site.
• Full cycle: about 50 swabs for 8 weeks active and about 75 for 12 weeks active; one 100 count box covers either.

Important Notes

Practical considerations for consistency and safety.

• Source pre-buffered NAD+ when possible. NAD+ stings primarily because it is acidic, near pH 3.5 to 4.0, and buffered formulations are pH adjusted at the manufacturer, which reduces burn meaningfully. Bicarbonate and other buffers are never mixed into the syringe.
• IM tolerates higher doses than sub-Q. IM is the default for doses above 100 mg, while sub-Q is reserved for smaller doses of 25 to 100 mg split across multiple injections.
• Comfort and site care: warm the vial about 10 minutes after refrigeration, distribute 1 mL across 30 to 60 seconds rather than pushing fast, split doses above 150 mg across two sites, and rotate sites systematically rather than injecting the same spot repeatedly. Persistent lumps lasting more than 7 days, spreading redness, fever, or breathing difficulty warrant medical evaluation.
• Stacking partners: SS-31 and MOTS-c for mitochondrial coverage in the Mito Stack; 5-Amino-1MQ to preserve NAD+ in adipose tissue by inhibiting NNMT; GLP-1 agonists for metabolic support during fat loss; injectable L-carnitine for fatty acid transport during oxidation.
• Contraindications: active cancer, since NAD+ supports cellular metabolism in tumors as well as healthy cells; pregnancy or breastfeeding, where data are insufficient; and severe hepatic or renal impairment. A history of cancer, cardiac arrhythmia, or chemotherapy calls for physician clearance.
• Regulatory Note: NAD+ injectable material is supplied for research use only and is not for human consumption; nothing described here is an approved medical use.

How This Works

NAD+ functions as the cellular redox currency spent across four competing systems. First, mitochondrial electron transport: NAD+ accepts electrons from glycolysis and the TCA cycle to drive ATP synthesis. Second, DNA repair: PARP enzymes consume NAD+ to repair single strand and double strand breaks, and chronic stress depletes pools through PARP overactivation. Third, sirtuin activity: SIRT1 through SIRT7 are NAD+ dependent deacetylases that regulate metabolism, circadian rhythm, and mitochondrial biogenesis via SIRT3 and PGC-1 alpha, and restrain NF-kB inflammatory signaling. Fourth, CD38 immune signaling: macrophages express CD38 NADase activity, which accelerates depletion under inflammation. Age related decline follows a self reinforcing loop in which senescent cells accumulate, secrete inflammatory signals (SASP), trigger CD38 upregulation of 200 to 300%, drain NAD+ faster than salvage can replace it, and disable the sirtuin brakes on inflammation; SARS-CoV-2 and other viral infections trigger this same metabolic collapse acutely. Exogenous NAD+ administration raises tissue pools and restores the enabling conditions for these systems to function, and clinical research has demonstrated brain NAD+ elevation in the NADPARK trial via MRS, blood pressure reduction of 5 to 10 mmHg with NR at 1,000 mg per day, and insulin sensitivity improvements with NMN at 250 mg per day in prediabetic women.

Benefits & Side Effects

Observations from the published record.

• Cardiovascular: NR at 1,000 mg per day reduced systolic blood pressure 5 to 10 mmHg in older adults, with improved arterial dilation and endothelial nitric oxide via SIRT1 activation.
• Metabolic and cognitive: NMN at 250 mg per day improved insulin sensitivity in prediabetic women and supports redox capacity during GLP-1 fat oxidation, while the NADPARK trial demonstrated that oral NR raises brain NAD+ measurable by MRS, with clinical benefits in Parkinson's patients under ongoing replication.
• Inflammation and recovery: NR reduced IL-6 and TNF alpha in older adults within weeks through restoration of the sirtuin and NF-kB axis, and subjective energy improvements are reported within the first week of IV or IM loading, building through weeks 1 to 4.
• Long COVID: a pilot trial (n=36) combining NAD+ patches with low dose naltrexone reported a 52% responder rate; SARS-CoV-2 directly suppresses NAMPT and NMNAT salvage enzymes while inducing PARP.
• Injection site reactions: burning, welting, and transient redness are universal because of pH. Buffered product, the IM route, slow injection, and warmed solution all minimize them. Infusing too quickly by IV can produce nausea, chest tightness, flushing, and lightheadedness, which resolve when the rate is slowed.
• Safety: oral NMN and NR were well tolerated at up to 2 g per day in trials, with no serious adverse events in published trials. Long term safety beyond 12 weeks remains limited.

Lifestyle Factors

Complementary strategies for best outcomes.

• Exercise upregulates NAMPT, the NAD+ salvage enzyme, by 12 to 30% in muscle; combining resistance and aerobic training covers both stimuli.
• Sleep of 7 to 9 hours protects the circadian oscillation of NAD+ recycling through NAMPT regulation, while disrupted sleep flattens it.
• Fasting activates salvage pathways and sirtuin signaling, and sauna may raise NAD+ about 20% acutely.
• Nutrition: niacin rich foods (chicken, tuna, mushrooms, peanuts) and tryptophan sources (turkey, eggs, dairy) support synthesis, apigenin and quercetin inhibit CD38 in cell studies with limited human data, and alcohol is limited because ethanol metabolism consumes NAD+ acutely.

Injection Technique

General subcutaneous guidance from clinical best-practice resources.

1. Wash hands thoroughly with soap and water.
2. Clean the vial stopper with an alcohol swab and allow to air-dry.
3. Select injection site (abdomen at least 2 inches from navel, outer thigh, or back of upper arm); clean and allow to dry.
4. Use a fine, short needle: 28 to 31 gauge insulin syringe; 5/16 to 1/2 inch (8 to 12 mm) length.
5. Pinch a fold of skin; insert the needle at approximately 45 degrees into the pinched subcutaneous layer.
6. Do not aspirate for subcutaneous injections. Depress the plunger slowly over 5 to 10 seconds.
7. Wait a few seconds before withdrawing the needle at the same angle.
8. Apply gentle pressure with a clean swab; do not rub vigorously.
9. Dispose of the used syringe immediately in a sharps container. 10 For doses above 150 mg, consider splitting into two injections at different sites. .

References

Source citations for further reading.

1. Covarrubias AJ et al. NAD+ metabolism and its roles in cellular processes during ageing (Nat Rev Mol Cell Biol, 2021).
2. Camacho-Pereira J et al. CD38 dictates age-related NAD decline and mitochondrial dysfunction (Cell Metabolism, 2016).
3. Grant R et al. Pharmacokinetics of intravenous NAD+ in humans (Redox Biology, 2019).
4. Brakedal B et al. NR increases brain NAD+ in Parkinson's disease, NADPARK trial (Nature Communications, 2024).
5. Martens CR et al. Chronic NR supplementation elevates NAD+ and reduces systolic BP in older adults (Nature Communications, 2018).
6. Yoshino M et al. NMN increases muscle insulin sensitivity in prediabetic women (Science, 2021).