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Researcher adjusting an electrophysiology recording stage
Neuro · Laboratory protocol

Semax

Heptapeptide ACTH(4-10) analog with a C-terminal Pro-Gly-Pro extension. Studied in BDNF/TrkB and neurotrophic signalling models.

For laboratory research use only. Material is supplied as a research reference standard and is not approved by Health Canada. Protocols below are compiled from published literature and supplier documentation; they are reference information, not medical advice. Anyone with a health condition or on medication should speak to a physician first.

Before you begin

  • • Lot-matched certificate and laboratory notebook
  • • Calibrated pipettes, low-binding tubes and suitable PPE
  • • Validated solvent, assay reagents and model-specific SOP
  • • Appropriate positive, negative and vehicle controls

Workflow

  1. 1Review the lot-matched COA and record identity, purity, batch number and received condition.
  2. 2Let the sealed vial equilibrate to room temperature before opening to limit condensation.
  3. 3Calculate the target stock concentration with C = mass / volume. Use only a solvent validated for the planned assay.
  4. 4Add solvent slowly down the vial wall, mix gently and avoid vortexing unless the validated method permits it.
  5. 5Prepare a log-spaced pilot concentration series; determine the usable range empirically instead of assuming a biological dose.
  6. 6Run the defined controls and technical replicates alongside every plate or experiment.
  7. 7Record preparation time, temperature, equipment, deviations and raw-data file locations.

Assay design

Neurotrophic endpoints such as BDNF or TrkB expression, with collection times and normalization method fixed before the material is prepared.

Required controls

  • Vehicle-only negative control
  • Untreated baseline
  • Validated pathway control
  • Parallel viability measurement

Reported protocol

Standard
300 mcg to 600 mcg on waking
Route
Intranasal or subcutaneous
Cycle length
14 to 21 days on, followed by 7 to 14 days off

Diluent

Sterile bacteriostatic water (0.9% benzyl alcohol) for multi-withdrawal stock, or sterile water for injection where a preservative-free stock is required by the assay.

Reconstitution concentrations

Solvent volume sets the stock concentration for this 10 mg vial. Lower volume gives a more concentrated stock and smaller working aliquots; higher volume gives a more dilute stock that is easier to measure accurately.

Stock concentration by solvent volume for a 10 mg vial
Solvent addedStock concentrationPer 0.1 mLPer unit (U-100)
1 mL10 mg/mL1 mg100 mcg
2 mL5 mg/mL500 mcg50 mcg
3 mL3.33 mg/mL333.33 mcg33.33 mcg

Working range

Neuronal culture work commonly runs the nanomolar to low-micromolar band.

Neurotrophic endpoints such as BDNF or TrkB expression, with collection times and normalization method fixed before the material is prepared.

Stability

Lyophilized
Stable sealed and dry at the certificate temperature.
In solution
Establish empirically for your solvent and container.

Handling notes

  • Fix the collection timepoints in advance; transcriptional readouts are highly time-sensitive.
  • If comparing against Selank, run both concurrently. They are different molecules with different proposed mechanisms.
  • Use low-binding tubes for dilute working stocks.

Documentation checklist

Record lot and COA, mass balance, solvent and concentration, preparation timestamp, storage history, plate map, replicate plan, instrument settings, exclusions and deviations. Preserve raw data and analysis code with the experiment record.