Semax Reconstitution & Dosage Protocol
ACTH(4-7)PGP — Neuroprotective Heptapeptide — Complete laboratory preparation guide
Ares Research Lab Team
Analytical Chemistry Division
READ TIME
6 MIN READ
Laboratory reconstitution protocol for Semax heptapeptide. Covers sterile saline preparation, rapid dissolution, oxidation protection, and research dosage ranges for neuronal culture and neuroprotective studies.
CAS NUMBER
80714-61-0
MOLECULAR FORMULA
C33H43N9O8S
MOLECULAR WEIGHT
708.82 g/mol
PURITY
≥ 99.5% (HPLC)
SEQUENCE
Met-Glu-His-Phe-Pro-Gly-Pro
RECONSTITUTION PROCEDURE
Semax is highly hygroscopic and water-soluble. Reconstitution is rapid, but the N-terminal methionine residue is susceptible to oxidation. Work quickly once the vial seal is broken and protect from light throughout handling.
Equilibrate and Inspect
Allow vial to reach ambient temperature for 10 minutes. Semax is hygroscopic — work quickly once opened. Inspect for uniform white powder; brown discoloration indicates oxidation.
Draw Sterile Saline
Use sterile saline (0.9% NaCl) or bacteriostatic water. For 10mg vial with target 5 mg/ml: draw 2.0ml. Semax dissolves rapidly due to its small molecular weight.
Inject and Dissolve
Inject solvent slowly down the inner vial wall. Semax dissolves almost instantly — gently invert 3–5 times. Do not shake or vortex.
Verify Clarity
Solution must be completely clear and colorless. Brown discoloration suggests methionine oxidation — discard immediately if observed.
Semax is exceptionally soluble but reconstituted solution is stable for only 7 days at 4°C. Aliquot immediately into single-use portions and freeze at -20°C for maximum stability. Protect from light at all stages.
CONCENTRATION & DOSAGE REFERENCE
Standard Ares Research Semax vials contain 10mg of lyophilized heptapeptide. The table below shows concentration after reconstitution.
RESEARCH DOSAGE RANGES
- →Neuronal cell culture (primary cortical/hippocampal): 10–100 mcg/ml in neurobasal/B27 media, 24–72 hours
- →Acute synaptic plasticity (LTP/LTD electrophysiology): 1–10 mcg/ml bath application for 30–60 minutes
- →Neuroprotection assays (OGD/reperfusion): 10–50 mcg/ml added 1 hour before insult
- →BDNF/NGF expression studies: 10–100 mcg/ml, qPCR/Western blot readout at 24–72 hours
STORAGE & STABILITY
Semax stability is limited by methionine oxidation and hygroscopicity. Strict light protection and cold-chain handling are essential.
- →Lyophilized powder: stable ≥ 24 months at -20°C, sealed and protected from light and moisture
- →Reconstituted solution: 7 days at 2–8°C in amber vial; aliquot and freeze at -20°C within 24 hours for long-term studies
- →Aliquots at -20°C: stable ≥ 3 months; thaw once and discard unused portion
- →Protect from UV, fluorescent light, and direct sunlight at all stages
- →Avoid repeated freeze-thaw cycles; Met⁹ oxidation accelerates with each cycle
pH & COMPATIBILITY NOTES
Semax is stable in neutral to mildly acidic pH. Avoid alkaline buffers and oxidizing conditions.
- →Optimal pH range: 6.0–7.5 in reconstituted solution
- →Do not expose to alkaline buffers (pH > 8.0) — accelerates degradation
- →Avoid oxidizing agents (H₂O₂, bleach, strong acids) — oxidizes Met⁹ residue
- →Compatible with standard neurobasal, DMEM/F12, and PBS buffers
- →Do not mix with reducing agents (ascorbic acid, glutathione) in the same stock vial
ANALYTICAL SPECIFICATIONS
REFERENCES & CITATIONS
1. Medvedeva, E.V. et al. (2017). Semax and its effects on BDNF expression. Bulletin of Experimental Biology and Medicine, 163(3), 339–342.
2. Koroleva, S.V. et al. (2015). Neuroprotective effects of Semax in cerebral ischemia. Neuroscience and Behavioral Physiology, 45(6), 678–684.
3. Guseva, M.V. et al. (2010). Effect of Semax on neuronal survival. Cellular and Molecular Neurobiology, 30(3), 381–389.
RESEARCH DISCLAIMER
All protocols are intended solely for qualified laboratory research professionals. Not for human use or consumption. For in-vitro and laboratory research only.
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— FREQUENTLY ASKED QUESTIONS
Q: What solvent should I use for Semax reconstitution?
Use sterile saline (0.9% NaCl) or bacteriostatic water. Both dissolve Semax instantly. Saline is preferred for neuronal culture applications to maintain isotonicity. Bacteriostatic water is acceptable for stock preparation if antimicrobial preservation is needed.
Q: How long does reconstituted Semax last?
Reconstituted Semax is stable for approximately 7 days at 2–8°C when protected from light. For studies extending beyond 7 days, aliquot immediately after reconstitution into single-use portions and freeze at -20°C. Thaw each aliquot once and discard unused portion.
Q: Why did my Semax solution turn brown?
Brown discoloration indicates oxidation of the N-terminal methionine residue. This is accelerated by light exposure, alkaline pH, and oxidizing agents. Discard brown or yellow solutions — oxidized Semax has significantly reduced biological activity in MC4R and BDNF assays.
Q: What concentration should I use for BDNF expression studies?
For primary cortical or hippocampal neuron cultures, Semax is typically evaluated at 10–100 mcg/ml in neurobasal media with B27 supplement. BDNF and NGF expression are assessed by qPCR, Western blot, or ELISA at 24–72 hours post-treatment. Include a vehicle control with media alone.
Q: Can Semax be used in electrophysiology experiments?
Yes. Semax is commonly used in acute slice electrophysiology for LTP/LTD studies. Bath apply at 1–10 mcg/ml for 30–60 minutes during recording. The peptide is stable in ACSF for 2–4 hours at room temperature. Prepare fresh working solution for each recording session.
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