📄 NovaVitality Peptides — Research Use Only (RUO)
Document ID: NV-HSP-MOTSC-2025v1
Effective Date: 09 December 2025
Classification: Internal Research Guidance — Not for Clinical or Regulatory Submission

🧪 MOTS-c (Mitochondrial ORF-derived Peptide, 16-aa)

Handling, Reconstitution & Storage Protocol

For In Vitro and Preclinical Laboratory Research Use Only

⚠️ WARNING: This protocol applies exclusively to research-grade MOTS-c (Catalogue: NV-MOTSC-10). This material is NOT for human or animal administration. Misuse violates the UK Human Medicines Regulations 2012, FDA 21 CFR §1271, and EU Directive 2001/83/EC.

1. Product Overview

Parameter	Value
Peptide Name	MOTS-c (Mitochondrial Open Reading Frame of the 12S rRNA-c)
Sequence	MRWQENQARGGLGLLG (N-terminal methionine included; amidated C-terminus standard)
Purity (HPLC)	≥98% (typical batch: 98.5–99.4%)
Molecular Weight	1724.03 g/mol (monoisotopic)
Appearance	White to off-white lyophilized powder
Solubility	Moderate in acidic aqueous buffers (e.g., 0.1% acetic acid in H₂O or 0.9% NaCl); poor in neutral pH water

2. Pre-Handling Safety & Facility Requirements

✅ Required Before Use:

Work within a certified Class II Biological Safety Cabinet (BSC) or laminar flow hood (ISO 5 environment).
Wear appropriate PPE: lab coat, nitrile gloves, safety goggles.
Ensure access to spill containment kit (absorbent pads, 70% ethanol, biohazard bag).
Confirm institutional approval for peptide handling (e.g., local Chemical Hygiene Plan compliance).

🚫 Prohibited:

Open vials outside controlled environments.
Use in non-sterile or non-research contexts (e.g., personal supplementation).
Store near volatile organics, strong oxidizers, or sources of UV light.

3. Reconstitution Procedure

3.1 Recommended Solvents

Solvent	Use Case	Notes
0.1% Acetic Acid in Sterile Water	Standard for cell culture assays	Optimal solubility; pH ≈ 3.5–4.0
Bacteriostatic Water (0.9% NaCl + 0.9% Benzyl Alcohol)	Short-term in vitro work (<72h)	Preservative extends shelf-life at 4°C; avoid if benzyl alcohol interferes with assay
DMSO (≤10% final conc.)	Stock for high-concentration aliquots	Use only for intermediate stocks; dilute ≥100-fold into aqueous buffer before biological use

📌 Tip: MOTS-c may form transient opalescence upon reconstitution — allow 5–10 min at RT with gentle swirling (no vortexing). Sonication (ice bath, 10–15 sec bursts) may aid dissolution.

3.2 Step-by-Step Reconstitution (10mg vial → 1 mg/mL stock)

Centrifuge lyophilized vial briefly (5 sec) to collect powder.
Wipe rubber stopper with 70% ethanol; allow to dry.
Using a sterile syringe, add 10 mL of chosen solvent slowly down the vial wall.
Gently rotate vial for 2–3 min until fully dissolved (no visible particulates).
Aliquot immediately into sterile, low-binding microtubes (e.g., 100 µL × 100 tubes).
Flash-freeze in liquid N₂ or dry ice/ethanol bath.
Store per Section 4.

❗ Do not reconstitute entire 10mg vial if single-use doses are smaller — partial reconstitution minimises degradation.

4. Storage Recommendations

Form	Temperature	Duration	Stability Notes
Lyophilized (unopened)	−20°C, desiccated, dark	≥24 months	Keep in original amber vial with desiccant
Lyophilized (opened)	−20°C, sealed with parafilm + desiccant	≤6 months	Minimise air exposure; purge with argon if possible
Reconstituted (aliquoted, frozen)	−80°C	≥12 months	Avoid frost-free freezers (temperature cycling degrades peptide)
Reconstituted (refrigerated)	2–8°C	≤7 days	For bacteriostatic water solutions only; monitor for turbidity

🔬 Stability Monitoring:

Visual check: Discard if cloudiness, precipitation, or colour change (yellow/brown) occurs.

For critical assays: Validate activity via pilot dose–response (e.g., AMPK phosphorylation in C2C12 myotubes) if stored >6 months.

5. Handling in Experimental Workflows

Cell Culture: Filter-sterilize (0.22 µm PVDF) after reconstitution if not using pre-sterile solvents.
In Vivo Rodent Studies: Only use under approved IACUC protocol. Inject within 2h of thawing; keep on ice.
Analytical Work (HPLC/MS): Dilute in 0.1% FA in 50:50 ACN:H₂O for optimal peak shape. Avoid phosphate buffers (ion suppression).

6. Disposal

Dispose of unused peptide and contaminated materials as chemical-biological mixed waste, per local regulations (e.g., UK: COSHH-compliant hazardous waste stream). Do not pour down the sink.

7. Certificate of Analysis (CoA) & QC

Each batch includes:

HPLC Chromatogram (C18, gradient elution)
MALDI-TOF or ESI-MS mass confirmation
Endotoxin level (typical: <1.0 EU/mg — not sterile or pyrogen-free)
Water content (KF titration)

📥 Request your batch-specific CoA by emailing [email protected] with your order number and catalogue ID.

8. References (Selected Research)

Lee et al. (2015). Cell Metabolism 21(3):441–448. (Original MOTS-c discovery)
Reynolds et al. (2021). Nature Communications 12:3895. (Mechanism: folate-AMPK axis)
Ming et al. (2023). Aging Cell 22(4):e13859. (Age-related metabolic rescue in mice)

📚 For protocol citations, please reference NovaVitality (2025). MOTS-c Handling & Storage Protocol v1.

✅ Document Prepared By: NovaVitality Ltd
📧 Contact: [email protected] | 🌐 novavitality.co.uk