Recombinant Mouse/Human TGF-β3

Recombinant TGF-β3 is a versatile and potent research tool with applications spanning developmental biology, tissue engineering, wound healing, and immunology. Here are the key reasons to incorporate it into your research:

Developmental and Tissue Regeneration Applications

TGF-β3 plays critical roles in embryonic development and tissue homeostasis that make it invaluable for regenerative medicine research. The protein is essential for palate development, and its absence causes cleft palate formation. Beyond craniofacial development, TGF-β3 regulates lung development and controls fundamental processes like cell adhesion, migration, and differentiation. This makes it particularly useful for studying developmental mechanisms and for tissue engineering applications targeting bone, cartilage, and other tissues.

In bone regeneration studies, TGF-β3 promotes osteogenic differentiation of stem cells and enhances osteoblast activity. Research demonstrates that TGF-β3-loaded scaffolds significantly increase expression of osteogenic markers including Runx2, BMP-2, and collagen I, making it suitable for cranial defect repair and orthopedic applications.

Wound Healing and Tissue Repair

TGF-β3 accelerates wound repair through multiple mechanisms. The protein regulates cell adhesion and movement in wounded tissues, functioning as a molecular "traffic control" system that coordinates dermal and epidermal cell migration during the healing process. In fascial wound healing models, TGF-β3 treatment increases breaking strength, collagen deposition, and cellular proliferation.

Clinical evidence supports therapeutic potential: TGF-β3 treatment has demonstrated efficacy in improving healing of pressure ulcers and reducing the severity and duration of chemotherapy-induced oral mucositis. The protein also stimulates scar-free healing and enhances glycosaminoglycan production by mesenchymal stromal cells, making it valuable for studying mechanisms of regenerative versus fibrotic healing responses.

Cartilage and Joint Research

TGF-β3 is recognized as a potential therapeutic target for osteoarthritis due to its protective effects on cartilage. The protein drives R-Smad-dependent signaling pathways that increase expression of chondroitin sulfate synthase 1 (CHSY1), an enzyme essential for proteoglycan synthesis. This signaling involves both Smad2/3 phosphorylation and MAPK pathway activation (ERK, p38, and JNK), providing multiple research angles for understanding chondrogenesis and cartilage maintenance.

Immunological Research

TGF-β3 exhibits context-dependent immunomodulatory functions that differ from other TGF-β isoforms. The protein can regulate systemic autoimmune diseases by inhibiting B cell responses and promoting regulatory T cell function. Research indicates TGF-β3 has potential preventive roles in lupus and other autoimmune conditions, with therapeutic effects demonstrated in murine disease models. Additionally, TGF-β3 can induce cell proliferation and angiogenesis while selectively promoting or inhibiting specific immune events.

Technical Advantages

Recombinant TGF-β3 offers practical advantages for laboratory work. Commercial preparations typically achieve >95% purity with low endotoxin levels (≤1 EU/μg), ensuring reliable and reproducible results. The protein has a molecular weight of approximately 25.8 kDa and demonstrates high bioactivity, with effective dose-response curves in standard proliferation assays.

The protein is optimized for multiple experimental formats including ELISA, Western blotting, and inhibition assays, as well as cell culture and functional assays. Its versatility extends to stem cell differentiation studies and T-cell regulation applications.

Research Flexibility

The availability of both human and mouse variants allows species-appropriate experimental design. Whether conducting mechanistic studies in murine models or translational research with human cells, recombinant TGF-β3 provides a standardized, reproducible tool for investigating growth factor signaling, tissue regeneration, immune tolerance, and fibrosis.

Yes, recombinant Mouse/Human TGF-β3 can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system and matches the species and isoform specificity of your antibodies.

Key considerations and supporting details:

• Recombinant TGF-β3 proteins are commonly used as standards in ELISA assays for quantifying TGF-β3 in biological samples. Many commercial ELISA kits are calibrated using highly purified recombinant TGF-β3, and protocols specify the use of recombinant standards for generating standard curves.
• Human and mouse TGF-β3 are 100% sequence homologous, so a recombinant protein covering both species is suitable for assays targeting either species, assuming your antibodies recognize both forms.
• Standard curve generation: The standard (recombinant TGF-β3) is serially diluted to create a standard curve, which is then used to interpolate the concentration of TGF-β3 in unknown samples.
• Assay specificity: Ensure that the ELISA antibodies are specific to TGF-β3 and do not cross-react with other TGF-β isoforms. High-quality ELISA kits report minimal cross-reactivity and are validated for use with recombinant standards.
• Protein format: For ELISA calibration, use recombinant TGF-β3 that is carrier-free or formulated for immunoassay use. Some suppliers recommend using BSA-containing formulations for stability, but carrier-free proteins are preferred if you need to avoid potential interference.
• Validation: It is important to confirm that the recombinant standard behaves similarly to native TGF-β3 in your assay matrix. This is typically validated by the kit manufacturer, but if you are developing a custom assay, you should verify recovery and linearity with your samples.

Best practices:

• Use the recombinant TGF-β3 standard within the validated range of your ELISA kit’s standard curve.
• Prepare and store the standard according to the manufacturer’s instructions to maintain stability and activity.
• If you are using a custom or in-house ELISA, validate the recombinant standard for parallelism and recovery in your sample matrix.

Summary Table: Use of Recombinant TGF-β3 as ELISA Standard

In summary: Recombinant Mouse/Human TGF-β3 is appropriate for use as a standard in ELISA quantification, provided it matches your assay’s specificity and is validated for your application.

Recombinant Mouse/Human TGF-β3 has been validated for a wide range of applications in published research, including:

• Cell culture and differentiation studies: TGF-β3 is used to support cell culture and to induce differentiation, particularly in stem cell and chondrocyte differentiation protocols.
• Functional assays: It is employed in bioassays to measure biological activity, such as inhibition of IL-4-dependent proliferation of HT-2 mouse T cells.
• ELISA and Western Blotting: TGF-β3 is used as a standard or detection target in immunoassays like ELISA and Western Blot.
• Blocking and inhibition assays: It is utilized in assays to study receptor binding and signal transduction, including blocking assays to assess pathway inhibition.
• Wound healing and tissue repair: TGF-β3 is studied for its role in accelerating wound repair and regulating cell adhesion.
• Cartilage development and osteoarthritis research: TGF-β3 is investigated for its effects on chondrocyte viability, type II collagen production, and its potential as a therapeutic target in osteoarthritis.
• Stem cell pluripotency and organoid differentiation: TGF-β3 is used in studies of stem cell differentiation and organoid formation, particularly in cerebral and telencephalic organoid models.
• Cancer research: TGF-β3 is studied for its effects on cancer cell motility, metastatic potential, and tumor progression.
• Signal transduction studies: TGF-β3 is used to investigate Smad-dependent and -independent signaling pathways, including MAPK activation and transcription factor interactions.

These applications are supported by research in areas such as developmental biology, immunology, tissue engineering, and disease modeling.

To reconstitute and prepare Recombinant Mouse/Human TGF-β3 protein for cell culture experiments, dissolve the lyophilized protein in sterile 4 mM HCl to a concentration of 20–50 μg/mL, optionally including 0.1–1% carrier protein (such as BSA or HSA) to enhance stability and prevent adsorption to surfaces.

Detailed protocol:

• Centrifuge the vial briefly before opening to collect the lyophilized powder at the bottom.
• Add sterile 4 mM HCl directly to the vial to achieve your desired stock concentration (commonly 20–50 μg/mL).
  • For example, add 1 mL of 4 mM HCl to a 50 μg vial for a 50 μg/mL stock.
• Include carrier protein (optional but recommended for stability): Add 0.1–1% BSA or HSA to the reconstitution buffer, especially if you plan to store aliquots or use low working concentrations.
• Gently swirl or tap the vial to dissolve the protein. Avoid vigorous vortexing to prevent denaturation.
• Aliquot the reconstituted stock to minimize freeze-thaw cycles.
• Storage:
  • Store aliquots at 2–8 °C for up to 1 week.
  • For longer-term storage, freeze at –20 °C or below.
  • Avoid repeated freeze-thaw cycles to maintain bioactivity.

Dilution for cell culture:

• Before adding to cell culture, dilute the stock solution into your cell culture medium to the desired final concentration.
• If the stock contains HCl, ensure the final HCl concentration in the culture medium is negligible and does not affect cell viability.
• If using carrier protein, maintain at least 0.1% BSA/HSA in working solutions to prevent protein loss due to adsorption.

Notes:

• Some protocols allow reconstitution in sterile water at 0.1 mg/mL, but 4 mM HCl is preferred for optimal solubility and stability for TGF-β3.
• Always consult the specific product datasheet for any manufacturer-specific recommendations.

Summary Table:

This protocol is suitable for both mouse and human recombinant TGF-β3 due to their high sequence homology and similar biochemical properties.