Recombinant Human Decorin, CF

Recombinant human decorin, CF (carrier-free) offers several compelling advantages for research applications, particularly in studies involving extracellular matrix biology, fibrosis, cancer research, and tissue engineering.

Key Biological Functions

Decorin is a small leucine-rich proteoglycan (SLRP) with multifaceted biological activities that make it valuable for research. The protein functions primarily by inhibiting transforming growth factor-β (TGF-β), a major driver of fibrotic responses across multiple tissues. This antifibrotic mechanism has demonstrated efficacy in cardiac, ocular, cutaneous, hepatic, and muscular tissues.

Beyond its antifibrotic properties, decorin modulates collagen fibrillogenesis, regulates extracellular matrix (ECM) integrity, and influences cell proliferation and signaling pathways. The protein also demonstrates anti-tumorigenic properties by suppressing cancer cell growth and metastasis through attenuation of EGFR-mediated signaling and induction of apoptosis.

Research Applications

Cancer and Tumor Biology – Decorin suppresses tumor progression, inhibits invasion, and reduces metastatic potential across various cancer types. Researchers can use it to study tumor microenvironment dynamics and screen potential anti-cancer therapeutics.

Tissue Engineering and Regenerative Medicine – As a recombinant growth factor and signaling protein, decorin supports stem cell differentiation and tissue development, making it essential for developing cell therapies and tissue engineering approaches.

Fibrosis Research – The protein's potent antifibrotic activity makes it invaluable for investigating pathological fibrosis in multiple organ systems and evaluating therapeutic interventions.

Cell Culture and Bioassays – Decorin is widely used in whole-cell bioassays and cell culture applications to study cellular behavior and optimize experimental models.

Technical Advantages

The carrier-free formulation provides high purity (>95%) with minimal endotoxin contamination (<0.1 EU/µg), ensuring reliable and reproducible results. The protein is produced in insect cell systems and demonstrates consistent biological activity validated through collagen fibrillogenesis assays. These specifications make it suitable for sensitive applications including mass cytometry, Western blotting, and protein arrays.

Recombinant Human Decorin, Carrier Free (CF), can be used as a standard for quantification or calibration in ELISA assays, provided it is highly purified and its concentration is accurately determined. This approach is widely accepted in ELISA methodology for quantifying target proteins.

Key considerations and best practices:

• Purity and Concentration: The recombinant protein should be highly purified, and its concentration must be precisely known. Carrier-free (CF) preparations are suitable when you want to avoid interference from stabilizers like BSA, which can affect assay specificity or downstream applications.
• Standard Curve Preparation: Prepare a serial dilution of the recombinant Decorin in the same buffer or diluent used for your samples to generate a standard curve. This allows for accurate quantification of Decorin in unknown samples.
• Validation: Confirm that the recombinant Decorin standard is recognized by the antibodies used in your ELISA. Most commercial Decorin ELISA kits use recombinant human Decorin as their calibration standard, demonstrating its suitability for this purpose.
• Carrier-Free vs. Carrier-Added: Carrier-free protein is recommended when BSA or other carriers could interfere with your assay. If stability is a concern, carrier-added versions may be preferable, but only if BSA does not interfere with your detection system.
• Documentation: Record the lot number, concentration, and preparation details for reproducibility and traceability.

Limitations and additional notes:

• If your ELISA is designed to detect native Decorin only (e.g., specific conformational epitopes), confirm that the recombinant standard is suitable for calibration.
• Always follow the specific instructions provided with your ELISA kit regarding standard preparation, as requirements may vary depending on antibody specificity and assay format.

Summary Table: Recombinant Decorin as ELISA Standard

In conclusion, recombinant human Decorin (CF) is appropriate for use as a standard in ELISA quantification, provided it meets the above criteria and is compatible with your assay system.

Recombinant Human Decorin, CF has been validated in published research for a range of applications, primarily centered on its biological activity in modulating cell signaling, fibrosis, tumorigenesis, and tissue remodeling. The most commonly validated applications include:

• Bioassays: Used to assess decorin’s effects on cell proliferation, apoptosis, migration, and signaling pathways, particularly in the context of cancer, fibrosis, and tissue repair.
• Cell Culture: Applied to cultured cells to study its impact on cellular processes such as apoptosis, differentiation, and response to growth factors.
• In Vivo Studies: Administered in animal models to evaluate therapeutic effects, such as reducing fibrosis, modulating inflammation, and inhibiting tumor growth.
• Control Protein: Used as a control in experiments investigating extracellular matrix interactions and signaling.
• ELISA (Standard): Utilized as a standard in ELISA assays for quantifying decorin or related analytes.

Key validated research applications and contexts:

• Antifibrotic studies: Decorin has been validated for inhibiting TGF-β signaling and reducing fibrosis in tissues such as heart, liver, skin, and muscle, both in vitro and in vivo.
• Cancer research: Used to suppress tumorigenesis, invasion, and metastasis in various cancer models, including breast cancer and glioma, by antagonizing receptor tyrosine kinases (e.g., EGFR) and modulating the tumor microenvironment.
• Wound healing and tissue remodeling: Applied in models of wound healing and tissue repair to study its effects on extracellular matrix organization and cellular responses.
• Ophthalmology: Investigated for its role in inhibiting lens epithelial cell apoptosis and in models of age-related macular degeneration.
• Cardiovascular and metabolic disease: Used in studies of diabetic cardiomyopathy and other metabolic disorders to assess its anti-inflammatory and antifibrotic effects.
• Immunomodulation: Explored for its ability to regulate immune cell migration and function, particularly in reproductive and inflammatory contexts.

Summary Table of Validated Applications

Additional notes:

• Decorin’s validated use is broad, spanning both basic mechanistic studies and preclinical therapeutic models.
• Most studies employ concentrations and protocols optimized for their specific system; users should consult primary literature for detailed methods.

If you require protocols or more detailed application notes for a specific research context, please specify the intended use.

To reconstitute and prepare Recombinant Human Decorin, CF protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute the protein in sterile phosphate-buffered saline (PBS) or sterile distilled water to a concentration of 0.1–0.5 mg/mL (100–500 μg/mL) as recommended by most protocols. For example, add 1 mL of buffer to 100 μg of protein for a 0.1 mg/mL solution.
• Gently mix by swirling or inverting; avoid vigorous shaking or vortexing to prevent protein denaturation.
• Allow the protein to dissolve at room temperature for 15–30 minutes. If undissolved material remains, gently mix for up to 2 hours at room temperature.
• If required for your application, adjust the pH to neutrality (pH 7.0–7.4) after reconstitution, especially if the protein was lyophilized from an acidic buffer.
• Filter sterilize the solution using a 0.2 μm filter if sterility is required for cell culture.

Dilution and Storage:

• For working solutions, dilute the reconstituted protein in cell culture medium or buffer containing a carrier protein (e.g., 0.1% BSA or 10% FBS) to minimize adsorption and loss of activity.
• For short-term storage (up to 1 month), keep at 2–8 °C under sterile conditions.
• For long-term storage, aliquot and freeze at −20 °C to −70 °C. Avoid repeated freeze-thaw cycles.
• For serum-free applications, use a non-animal stabilizer such as trehalose instead of BSA or FBS.

Application in Cell Culture:

• Add the reconstituted Decorin directly to the cell culture medium at the desired final concentration, as determined by your experimental design or literature precedent.
• Typical working concentrations for functional assays range from 1–10 μg/mL, but optimization may be required for your specific cell type and assay.

Summary Table: Recombinant Human Decorin Reconstitution

Always consult the specific product datasheet or Certificate of Analysis for any manufacturer-specific recommendations, as formulations and optimal conditions may vary.