Researcher positioning a cell-culture dish beneath a laboratory microscope

Healing · Laboratory protocol

KPV

Lys-Pro-Val, the C-terminal tripeptide of alpha-MSH. Studied for anti-inflammatory signalling independent of pigmentation pathways.

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

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

A stimulated inflammatory model (for example LPS-challenged cells) with NF-kB or cytokine readouts, and the parent alpha-MSH run alongside where the question is fragment-versus-parent activity.

Required controls

Vehicle-only negative control
Untreated time-zero control
Validated migration control
Parallel cytotoxicity or viability assay

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 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

Working range

Anti-inflammatory cell work commonly runs the micromolar band, higher than most peptides here because of the small size and the proposed intracellular route.

A stimulated inflammatory model (for example LPS-challenged cells) with NF-kB or cytokine readouts, and the parent alpha-MSH run alongside where the question is fragment-versus-parent activity.

Stability

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

Handling notes

Run alpha-MSH as the comparison arm when the research question is whether the fragment retains parent activity.
The stimulus control (stimulated, no peptide) matters more than the unstimulated baseline here.
Small peptides need proportionally higher concentrations; do not import a nanomolar range from receptor-agonist literature.

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.