Humanin Protocol Guide
Humanin is a 24-amino acid mitochondrial-derived peptide (MDP) encoded by the MT-RNR2 gene in mitochondrial DNA. Discovered through its ability to protect neurons from amyloid-beta toxicity associated with Alzheimer's disease, Humanin is now recognized as a key retrograde signaling molecule linking mitochondrial function to cellular survival. It activates the STAT3 pathway via the CNTFR/WSX-1/gp130 trimeric receptor, inhibits apoptosis through BAX interaction, improves insulin sensitivity via IGFBP-3 binding, and reduces inflammatory cytokines. The potent analog HNG (S14G-Humanin) is approximately 1000x more bioactive.
Protocol Overview
- Compound
- Humanin (HN) / HNG (S14G-Humanin analog)
- Category
- Mitochondrial-Derived Peptide (MDP)
- Mechanism
- STAT3 activation via trimeric receptor, BAX-mediated anti-apoptosis, IGFBP-3 binding for insulin sensitization, ROS reduction
- Sequence
- 24 amino acids (MAPRGFSCLLLLTSEIDLPVKRRA)
- Half-Life
- ~30 minutes (native); HNG analog has improved stability
- Vial Size
- 5mg lyophilized powder (typical)
- Route
- Subcutaneous injection
- Frequency
- Once daily
- Cycle Length
- 4-12 weeks
Dosing Protocol
| Protocol | Dose | Frequency | Route | Duration |
|---|---|---|---|---|
| HNG Conservative | 1-2 mcg | Once daily | Subcutaneous | 4-8 weeks |
| HNG Standard | 5-10 mcg | Once daily | Subcutaneous | 8-12 weeks |
| Native Humanin | 1-2 mg | Once daily | Subcutaneous | 4-8 weeks |
| Native Humanin High | 2-4 mg | Once daily | Subcutaneous | 4-8 weeks |
Key principle: HNG (S14G-Humanin) is the preferred research form due to 1000x greater potency. Native Humanin requires milligram dosing while HNG works in the microgram range. Administer on an empty stomach in the morning.
Reconstitution Instructions
Materials Needed
- Humanin or HNG lyophilized vial
- Bacteriostatic water (1-2mL)
- 3mL mixing syringe (18-20ga)
- Insulin syringes (29-31ga) for dosing
- Alcohol swabs
Reconstitution Steps (5mg HNG vial)
- Clean vial stopper with alcohol
- Draw 1mL bacteriostatic water
- Inject slowly down the vial wall
- Gently swirl (never shake)
- Wait 5 minutes for full dissolution
- Concentration: 5,000 mcg/mL
| Dose (HNG) | Volume (1mL recon) | Insulin Syringe |
|---|---|---|
| 1 mcg | 0.0002 mL | ~0.02 units (use serial dilution) |
| 5 mcg | 0.001 mL | 0.1 units (use serial dilution) |
| 10 mcg | 0.002 mL | 0.2 units (use serial dilution) |
| 50 mcg | 0.01 mL | 1 unit |
Note: Due to very small dosing volumes for HNG, serial dilution may be required. Reconstitute to a more dilute concentration (e.g., 50mcg/mL) for practical dosing accuracy.
Administration Guide
Injection Sites
- Primary: Abdomen (2 inches from navel)
- Alternate: Anterior thigh
- Alternate: Upper arm (deltoid region)
- Rotate injection sites daily
Timing & Storage
- Timing: Morning on empty stomach preferred
- Storage (lyophilized): -20°C for long-term, 2-8°C short-term
- Storage (reconstituted): Refrigerate 2-8°C, use within 14 days
- Protect from light and repeated freeze-thaw cycles
Expected Timeline
Side Effects & Monitoring
Common Side Effects
- Injection site irritation (mild)
- Mild headache (transient)
- Slight fatigue during initial days (uncommon)
Humanin has shown an excellent safety profile in preclinical studies. Endogenous levels decline naturally with age, and supplementation restores physiological signaling.
Precautions
- Limited human clinical trial data available
- May affect blood glucose - monitor if diabetic
- Not recommended during pregnancy or breastfeeding
- Theoretical concern with tumor cell cytoprotection - avoid with active malignancy
Stacking Recommendations
Compatible Compounds
- MOTS-c: Fellow MDP for complementary metabolic benefits
- SS-31: Cardiolipin stabilizer for synergistic mitochondrial support
- Selank: Anxiolytic peptide for cognitive optimization stack
- Epithalon: Telomerase activator for longevity protocol
- NAD+ precursors: Support mitochondrial NAD+ pool
Popular Stacks
- Longevity MDP Stack: Humanin + MOTS-c + SS-31
- Neuroprotection: Humanin + Selank + Semax
- Metabolic Health: Humanin + MOTS-c + NAD+
Blood Work Recommendations
| Panel | Markers | Timing |
|---|---|---|
| Basic Panel | CBC, CMP | Baseline, Week 6, Week 12 |
| Metabolic | Fasting glucose, insulin, HbA1c, HOMA-IR | Baseline, Week 6, Week 12 |
| Inflammatory | CRP, IL-6, TNF-alpha | Baseline, Week 6 |
| Oxidative Stress | 8-OHdG, F2-isoprostanes | Baseline, Week 8 |
| Cognitive (optional) | BDNF, NGF | Baseline, Week 8 |
Metabolic panels (fasting glucose, insulin, HOMA-IR) are the most practical tracking markers for Humanin protocols, given its well-documented insulin-sensitizing effects.
Related Tools & Resources
