Epithalon Reconstitution Protocol: Step-by-Step Guide for Laboratory Research Epithalon (Epitalon, Ala-Glu-Asp-Gly, MW 390.35 g/mol, CAS 307297-39-8) is a synthetic tetrapeptide derived from Epithalamin studied in cell-based research for telomerase activation, telomere dynamics, and pineal gland-associated regulatory pathway modulation. It is supplied as a lyophilised white powder in a 10mL sterile glass vial containing 10mg of peptide. As one of the smallest research peptides by molecular weight, epithalon is exceptionally water-soluble and reconstitutes instantly in BAC water. Materials Required Epithalon 10mg lyophilised vial Bacteriostatic water (BAC water) — 10mL vial 1mL or 2mL sterile syringe with needle Alcohol swabs (70% isopropyl) Nitrile gloves Permanent marker for labelling Refrigerator at 2–8°C for storage Pre-Reconstitution Preparation Inspect the epithalon vial to confirm the lyophilised powder appears white and intact with an unbroken seal. Remove from cold storage and equilibrate to room temperature for 10 minutes. Epithalon's tetrapeptide structure (only 4 amino acids, MW 390.35 g/mol) makes it the most water-soluble compound in the research peptide category — dissolution is essentially instantaneous upon contact with aqueous solvent. Step-by-Step Reconstitution Protocol Step 1 — Prepare workspace. Wipe work surface with 70% isopropyl alcohol and allow to dry. Wear nitrile gloves throughout. Step 2 — Swab both stoppers. Clean rubber stoppers on both vials with separate fresh alcohol swabs. Air dry 10 seconds each. Step 3 — Draw 2mL of BAC water. Draw 2mL of bacteriostatic water into a sterile syringe. For a 10mg epithalon vial, 2mL yields a 5mg/mL stock. A lower stock of 2.5mg/mL can be prepared using 4mL BAC water if preferred for your assay dilution scheme. Step 4 — Inject along the vial wall. Insert the needle into the epithalon vial and direct BAC water slowly down the inner glass wall over 10 seconds. Step 5 — Dissolve. Epithalon dissolves instantly — swirl gently for 10–15 seconds to confirm complete dissolution. The solution will be clear and colourless. Do not shake. Step 6 — Inspect and label. Confirm clarity. Label with reconstitution date, concentration, and initials. Resulting Concentration Adding 2mL BAC water to a 10mg epithalon vial yields a stock concentration of 5mg/mL (5000mcg/mL). For telomerase activation assays (TRAP assay) and pinealocyte cell model studies, working concentrations of 1ng/mL to 100mcg/mL are prepared by serial dilution into assay buffer or cell culture media. Concentration Reference Table BAC Water Volume AddedResulting Concentration 1mL10.0mg/mL 2mL5.0mg/mL (5000mcg/mL) 4mL2.5mg/mL (2500mcg/mL) Storage Conditions and Stability Store lyophilised epithalon at −20°C prior to reconstitution, protected from light and moisture. Once reconstituted, store at 2–8°C. Reconstituted epithalon in BAC water is stable for up to 28 days under refrigeration. Epithalon's small tetrapeptide structure is inherently stable — it is resistant to most endopeptidase cleavage due to its short length, and is stable across a wide pH range (3.0–10.0). Do not freeze reconstituted solution. For long-term aliquot storage, prepare single-use microcentrifuge aliquots and store at −80°C. Research Application Notes Epithalon is soluble in BAC water, sterile water, PBS, and essentially any aqueous buffer without co-solvent requirements. Its exceptional water solubility and small size mean it can be added directly to cell culture media at high concentrations without precipitation or cytotoxicity concerns at research concentrations. For telomerase research applications using the TRAP (telomeric repeat amplification protocol) assay, prepare working dilutions in the TRAP assay lysis buffer rather than cell culture media to ensure compatibility with PCR-based detection. Epithalon's mechanism of action in telomerase biology research involves interaction with DNA-associated regulatory elements — for chromatin immunoprecipitation (ChIP) or telomere length assays, verify no interference between epithalon and the assay detection chemistry before committing to full concentration-response protocols. Epithalon 10mg vials are available from Pinnacle Peptides, verified at ≥99% purity by HPLC and mass spectrometry. All content is intended for in vitro laboratory research purposes only. Not for human or animal consumption. Not intended to diagnose, treat, cure, or prevent any condition.