Recombinant Human Decorin

Recombinant human decorin (rhDecorin) offers substantial scientific value across multiple research applications due to its diverse biological functions and therapeutic potential.

Key Biological Functions

Decorin is a small leucine-rich proteoglycan with multifaceted roles in cellular and tissue biology. As a component of the extracellular matrix, it functions as a pan-receptor tyrosine kinase (RTK) inhibitor and modulates critical signaling pathways, particularly transforming growth factor-beta (TGF-β) signaling. This makes it particularly valuable for investigating fibrotic processes, inflammatory responses, and tumor biology.

Primary Research Applications

Anti-fibrotic Studies

Recombinant decorin demonstrates potent anti-fibrotic effects across multiple tissue contexts. It reduces myofibroblast-like gene expression and inhibits collagen fibrillogenesis, making it essential for research on wound healing, scar formation, and fibrotic disease models. Studies have shown efficacy in peritoneal dialysis-associated peritonitis, corneal fibrosis, and abdominal aortic aneurysm models.

Cancer Research

Decorin exhibits anti-tumor capabilities by suppressing cell growth, differentiation, survival, and metastasis. It has demonstrated efficacy in reducing tumorigenesis, invasion, and metastasis in inflammatory breast cancer models. Additionally, decorin enhances cancer immunotherapy by attenuating intratumoural TGF-β expression and increasing CD8+ T cell infiltration.

Cell Culture and Bioassay Applications

Recombinant decorin is widely used in cell culture systems and bioassays to investigate protein-protein interactions, signaling pathways, and cellular responses. It can be employed with or without carrier proteins (such as BSA) depending on your specific experimental requirements.

Neuroprotection and Inflammation

Topical decorin provides neuroprotective and anti-inflammatory effects, as demonstrated in corneal neuropathy models. This application extends to investigating inflammatory responses and tissue protection mechanisms.

Experimental Advantages

Using recombinant decorin provides standardized, reproducible results compared to native protein sources. The protein is available in multiple formulations—carrier-free or with stabilizing agents—allowing optimization for your specific experimental conditions. Its well-characterized biological activity makes it suitable for quantitative bioassays, ELISA standards, and mechanistic studies of TGF-β signaling and extracellular matrix biology.

Yes, recombinant human Decorin can generally be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately known. This practice is common in research ELISAs, where recombinant proteins are used to generate standard curves for quantifying the same protein in biological samples.

Key considerations and supporting details:

• ELISA Kits and Standards: Most commercial human Decorin ELISA kits use recombinant human Decorin as the standard for calibration. The standard curve generated with recombinant Decorin allows for the quantification of Decorin in serum, plasma, or cell culture supernatants.
• Specificity: These kits are validated to detect both natural and recombinant forms of human Decorin, ensuring that the recombinant standard is suitable for quantifying endogenous Decorin in biological samples.
• Purity and Quantification: The recombinant Decorin used as a standard should be highly purified and its concentration determined accurately (e.g., by absorbance at 280 nm or amino acid analysis). Impurities or inaccurate quantification can lead to errors in your standard curve and sample measurements.
• Formulation: If your recombinant Decorin contains carrier proteins (such as BSA), ensure this matches the formulation of the standards used in your ELISA kit, as carriers can affect assay performance.
• Validation: If you are developing your own ELISA or using a kit with a different standard, you should validate that your recombinant Decorin standard produces a linear, reproducible standard curve within the assay’s dynamic range.

Best Practices:

• Reconstitute and dilute the recombinant Decorin according to the ELISA kit’s instructions or your validated protocol.
• Prepare a serial dilution to generate a standard curve covering the expected concentration range of your samples.
• Confirm that the recombinant standard yields parallelism with endogenous Decorin in your sample matrix, indicating comparable immunoreactivity.

Summary Table: Use of Recombinant Human Decorin as ELISA Standard

In summary: You can use recombinant human Decorin as a standard for ELISA quantification, provided it is of high quality and validated for your assay system. Always follow best practices for standard preparation and validation to ensure accurate quantification.

Recombinant Human Decorin has been validated for a diverse range of applications in published research, primarily in the fields of cell biology, fibrosis, cancer, wound healing, and extracellular matrix modulation.

Validated Applications:

• Bioassays: Used to assess biological activity, such as modulation of cell proliferation, apoptosis, and differentiation in various cell types including fibroblasts, myoblasts, lens epithelial cells, and neural stem/progenitor cells.
• In Vivo Studies: Applied in animal models to investigate therapeutic effects on fibrosis, tumor growth, metastasis, ligamentum flavum hypertrophy, and wound healing.
• ELISA Standard: Utilized as a standard protein in ELISA assays for quantification and detection purposes.
• Control Protein: Employed as a control in experimental setups, such as wound healing models.
• Collagen Fibrillogenesis Assays: Used to study its ability to modulate collagen fibril formation, demonstrating delayed type I collagen fibrillogenesis at concentrations above 10 μg/ml.
• Mass Cytometry (CyTOF), Western Blotting, and Spatial Biology: Decorin has been used in advanced analytical techniques for protein detection and localization in tissue samples.
• Anti-Scarring and Anti-Fibrotic Therapy: Investigated as a topical agent to reduce scarring in skin diseases (e.g., dystrophic epidermolysis bullosa) and as an anti-fibrotic therapeutic in ocular and cardiovascular models.
• Cancer Research: Validated for suppression of tumorigenesis, invasion, metastasis, and lymphangiogenesis in breast cancer and other solid tumors.
• Immunomodulation: Studied for its role in modulating innate immunity and inflammatory responses via Toll-like receptor signaling.
• Glycoproteomics and Proteomics: Used in studies analyzing processed forms of decorin in human cardiac extracellular matrix.

Key Experimental Contexts:

• Cell Culture: Decorin is frequently used in cell culture systems to study its effects on cell behavior, ECM interactions, and signaling pathways.
• Animal Models: Validated in rodent models for in vivo efficacy in disease modification, including fibrosis, cancer, and vascular remodeling.
• Tissue and Organ Systems: Applied in studies of skin, heart, eye (trabecular meshwork), ligament, and tumor microenvironments.

Summary Table of Applications

Conclusion:
Recombinant Human Decorin is a versatile research tool validated for bioassays, in vivo disease models, ECM modulation, anti-fibrotic and anti-scarring therapies, cancer suppression, immunomodulation, and as a standard in analytical assays. Its applications span cell culture, animal models, and tissue-based studies, reflecting its broad utility in biomedical research.

To properly reconstitute and prepare Recombinant Human Decorin protein for cell culture experiments, follow these best practices based on manufacturer guidelines and scientific protocols:

1. Reconstitution Steps

• Centrifuge the vial briefly (e.g., 3000–3500 rpm for 5 minutes) before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute with sterile PBS or sterile distilled water as recommended by the product datasheet. Typical concentrations range from 0.1 to 1.0 mg/mL (e.g., 300 µg/mL for some products).
• Gently add buffer along the inner wall of the vial to minimize foaming.
• Allow the vial and buffer to equilibrate to room temperature before reconstitution.
• Gently mix by swirling or pipetting up and down. Do not vortex or shake vigorously, as this may denature the protein.
• Let the solution sit at room temperature for 15–30 minutes to ensure complete dissolution.

2. Dilution for Cell Culture

• Dilute the reconstituted protein to the desired working concentration using cell culture medium or PBS.
• Include carrier proteins (e.g., 0.1% BSA, 5% HSA, or 10% FBS) in the dilution buffer to stabilize the protein and prevent adsorption to surfaces.
• Avoid diluting with plain water for cell culture applications, as this may reduce protein stability.

3. Storage and Handling

• For short-term use (≤1 week): Store the reconstituted protein at 2–8°C.
• For long-term storage: Aliquot the protein and store at –20°C to –70°C in a manual defrost freezer.
• Avoid repeated freeze-thaw cycles, as each cycle can reduce protein activity.
• For carrier-free or animal-free applications: Use trehalose (e.g., 10.26%) as a stabilizer instead of BSA or FBS.

4. Application Notes for Cell Culture

• Decorin is typically used at concentrations above 10 µg/mL for biological activity (e.g., modulation of collagen fibrillogenesis).
• Decorin supports myoblast differentiation, angiogenesis, and can inhibit tumor progression in vitro.
• Ensure sterile technique throughout to prevent contamination.

Summary Protocol

1. Centrifuge vial briefly.
2. Reconstitute with sterile PBS or water to 0.1–1.0 mg/mL.
3. Gently mix and let sit at room temperature for 15–30 min.
4. Dilute to working concentration in medium or PBS with carrier protein.
5. Use immediately or aliquot and store at 2–8°C (≤1 week) or –20°C to –70°C (long-term).
6. Avoid vortexing and repeated freeze-thaw cycles.

Always refer to the specific product datasheet for exact reconstitution and storage instructions, as formulations may vary between suppliers.