KPV is a short tripeptide composed of the amino acids lysine (K), proline (P) and valine (V). It has gained attention in biomedical research for its anti-inflammatory, antimicrobial and tissue-regenerative properties. In many studies KPV has been shown to reduce cytokine production, inhibit neutrophil activation, and promote healing of skin and mucosal tissues. Because it is small, chemically stable, and inexpensive to synthesize, researchers often use KPV as a therapeutic agent in models of inflammatory bowel disease, periodontitis, wound healing, and even as an adjunct in vaccine formulations where modulation of the immune response is desirable.
KLOW Protocol
The KLOW protocol is a standardized method for evaluating the anti-inflammatory effects of KPV in vitro. The acronym stands for "Knock-down Lymphocyte-derived Oxidative-Stress-Related Wound-Healing." The steps are:
Cell Preparation – Primary human keratinocytes or murine macrophages are cultured to 80 % confluence in a 6-well plate.
Treatment – Cells receive KPV at concentrations ranging from 10 µM to 1000 µM for 24 h. A vehicle control (e.g., PBS) is included.
Stimulation – After pre-treatment, cells are stimulated with lipopolysaccharide (1 µg/mL) or TNF-α (20 ng/mL) to induce inflammation.
Readouts – Cytokine levels in the supernatant are measured by ELISA for IL-6, IL-8 and MCP-1. Reactive oxygen species production is quantified using a DCFDA fluorescence assay.
Data Analysis – Results are expressed as percent inhibition relative to the stimulated control. Statistical significance is assessed with one-way ANOVA followed by Tukey’s post-hoc test.
The KLOW protocol is widely adopted because it balances simplicity with reproducibility, allowing laboratories to compare their findings against a common benchmark.
Peptide Reconstitution Calculator
When working with peptide reagents like KPV, accurate reconstitution is critical. The Peptide Reconstitution Calculator helps researchers determine the correct volume of solvent needed to achieve a desired stock concentration. Users input:
Desired final concentration (µM or mg/mL)
Mass of peptide in milligrams
Molecular weight of the peptide
Volume of solvent to be used
The calculator outputs the exact amount of water, DMSO, or buffer required, along with the resulting molarity and a step-by-step reconstitution protocol. This tool is especially useful for peptides that are hygroscopic or prone to aggregation because it reduces errors that can arise from manual calculations.
Quick Reference
Below is a concise reference sheet summarizing key points about KPV:
Structure: Lysine-Proline-Valine
Molecular weight: 309.4 Da
Typical use concentrations: 10–1000 µM in cell culture; 1–10 mg/kg in animal models
- Skin wound healing assays - Oral mucosal inflammation models - In vitro macrophage activation studies - Adjunctive therapy in vaccine formulations to dampen excessive responses
Stability: Stable at room temperature for up to 48 h; freeze-thaw cycles should be limited
Storage: Dry powder stored at –20 °C; reconstituted solutions kept on ice and used within 24 h
By following the KLOW protocol, utilizing the Peptide Reconstitution Calculator for accurate dosing, and referring to this quick reference guide, researchers can effectively harness the therapeutic potential of the KPV peptide in a variety of experimental settings.
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