BPC-157
A 15-residue partial sequence (GEPPPGKPADDAGLV) of a protein found in gastric juice. Preclinical work centres on angiogenesis, fibroblast migration and the nitric-oxide pathway.
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
- 1Review the lot-matched COA and record identity, purity, batch number and received condition.
- 2Let the sealed vial equilibrate to room temperature before opening to limit condensation.
- 3Calculate the target stock concentration with C = mass / volume. Use only a solvent validated for the planned assay.
- 4Add solvent slowly down the vial wall, mix gently and avoid vortexing unless the validated method permits it.
- 5Prepare a log-spaced pilot concentration series; determine the usable range empirically instead of assuming a biological dose.
- 6Run the defined controls and technical replicates alongside every plate or experiment.
- 7Record preparation time, temperature, equipment, deviations and raw-data file locations.
Assay design
Scratch/wound-closure or transwell-migration assays with images captured at fixed intervals, normalized to viable cell count, with a parallel cytotoxicity plate so a migration effect is not confused with a viability effect.
Required controls
- Vehicle-only negative control
- Untreated time-zero control
- Validated migration control
- Parallel cytotoxicity or viability assay
Reported protocol
- Standard daily
- 250 mcg to 500 mcg per day
- Route
- Subcutaneous or intramuscular, near the affected area
- Cycle length
- 4 to 8 weeks continuous, followed by 2 to 4 weeks 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.
| Solvent added | Stock concentration | Per 0.1 mL | Per unit (U-100) |
|---|---|---|---|
| 1 mL | 10 mg/mL | 1 mg | 100 mcg |
| 2 mL | 5 mg/mL | 500 mcg | 50 mcg |
| 3 mL | 3.33 mg/mL | 333.33 mcg | 33.33 mcg |
| 5 mL | 2 mg/mL | 200 mcg | 20 mcg |
Working range
Cell-culture work commonly spans the nanomolar to low-micromolar band. Published rodent studies report effects across a very wide range, which is one reason the in-vitro literature is the more useful starting point for setting a series.
Scratch/wound-closure or transwell-migration assays with images captured at fixed intervals, normalized to viable cell count, with a parallel cytotoxicity plate so a migration effect is not confused with a viability effect.
Stability
- Lyophilized
- Notably robust as a dry cake. Keep sealed and dry at the certificate temperature.
- In solution
- Establish empirically. Acid stability in the dry state says nothing about stability in your assay buffer at 37 degrees Celsius.
Handling notes
- Time-zero images are the control that matters most in a closure assay. Capture them before anything is added.
- Normalize closure to viable cell count, not to field area alone.
- Run a parallel viability plate at every concentration in the series.
- The preclinical literature is broad and heterogeneous. Match your model to the specific paper you are testing against.
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.