Recombinant Human TGF-α

Recombinant Human TGF-α is widely used in research because it is a potent growth factor that regulates cell proliferation, differentiation, and tissue repair, making it essential for studies in cell biology, regenerative medicine, cancer, and tissue engineering.

Key scientific applications and advantages include:

• Cell Proliferation and Differentiation: TGF-α is a mitogen that activates epithelial cell proliferation, growth, and differentiation, especially in epidermal and epithelial cells. This makes it valuable for experiments requiring controlled stimulation of cell growth or modeling tissue development.

• Wound Healing and Tissue Engineering: TGF-α promotes tissue regeneration and accelerates healing, supporting its use in studies of wound repair and the development of engineered tissues.

• Cancer Research: TGF-α is involved in tumor growth, angiogenesis, and modification of the tumor microenvironment. It is frequently used to study cancer cell signaling, metastasis, and to screen potential anti-cancer therapies.

• Stem Cell and Regenerative Medicine: Recombinant TGF-α helps direct stem cell fate, supporting differentiation into specific tissues and enhancing protocols for cell therapy and tissue engineering.

• Bone Biology: TGF-α stimulates the formation of osteoclast-like cells and bone resorption, making it useful for investigating bone metabolism and diseases such as osteoporosis.

• Immunology and CNS Research: TGF-α modulates immune cell activity, reduces inflammation, and contributes to recovery in central nervous system injury models by decreasing infiltrating T cells and demyelination.

• Reliable Bioactivity: Recombinant production ensures high purity, batch-to-batch consistency, and biological activity, which are critical for reproducible experimental results.

• Mechanistic Studies: TGF-α binds to EGFR, triggering receptor dimerization and downstream signaling pathways (e.g., AKT, MAPK), allowing detailed analysis of cell signaling mechanisms.

In summary, using recombinant human TGF-α enables precise control over experimental conditions, supports a wide range of biomedical research applications, and provides a reliable tool for dissecting cellular and molecular mechanisms relevant to development, disease, and therapy.

You can use recombinant human TGF-α as a standard for quantification or calibration in ELISA assays, provided the ELISA kit is validated for both recombinant and natural forms of TGF-α. Many commercial ELISA kits for human TGF-α are designed to recognize both natural and recombinant proteins, and their standard curves are often generated using recombinant TGF-α.

Key considerations:

• Kit Validation: Confirm that your ELISA kit is validated for use with recombinant TGF-α. Many kits specify in their documentation that they detect both natural and recombinant forms, and that standard curves generated with recombinant protein are suitable for quantifying endogenous TGF-α in biological samples.
• Parallelism: Ideally, the standard curve generated with recombinant TGF-α should be parallel to the response obtained from endogenous (natural) TGF-α in your sample matrix. This ensures accurate quantification.
• Protein Source and Purity: Use a recombinant TGF-α standard that is well-characterized, with known concentration and purity, and preferably produced in a system that yields a protein similar in post-translational modifications to the native form (e.g., E. coli-expressed TGF-α is commonly used for standards).
• Documentation: Some manufacturers and protocols explicitly state that their kits are calibrated using recombinant human TGF-α and that results obtained with natural samples are comparable, as the standard curves are parallel.

Exceptions and Cautions:

• Bioassay Use: Recombinant proteins intended as ELISA standards are not always suitable for bioassays or functional studies, as they may not be tested for biological activity.
• Kit-Specific Requirements: Always check the specific ELISA kit instructions. Some kits may be optimized for native protein detection and may not guarantee equivalence with recombinant standards.

Summary Table: Recombinant TGF-α as ELISA Standard

Best Practice:
Always verify your specific ELISA kit’s documentation to ensure compatibility with recombinant TGF-α as a standard. If validated, recombinant human TGF-α is widely accepted and commonly used for calibration and quantification in ELISA assays for TGF-α.

Recombinant human TGF-α has been validated for multiple research applications across diverse experimental contexts. The primary validated applications include:

In Vitro Applications

Recombinant TGF-α is extensively used in functional assays, where it demonstrates dose-dependent bioactivity. The protein induces proliferation of BALB/3T3 cells with an ED₅₀ ranging from 0.02-0.1 ng/mL. This makes it particularly valuable for quantifying ligand-induced cellular responses and receptor activation studies.

The protein is also validated for ELISA applications, enabling quantitative detection and measurement of TGF-α in various sample matrices. Additionally, it serves as a tool in Western blot analyses and blocking assays, where it can be used to assess receptor-ligand interactions and signaling pathway inhibition.

Cell Culture and Bioassay Applications

TGF-α is employed in cell culture stimulation experiments to investigate growth factor-dependent cellular responses. Published research demonstrates its use in bioassay applications across multiple cell types and species, including human, mouse, rat, and hamster samples. These studies examine TGF-α's role in cell proliferation, migration, and signaling pathway activation in various cancer models and normal cell systems.

Therapeutic and Translational Research

Beyond basic research, recombinant TGF-α has been investigated for therapeutic applications including wound healing, tissue engineering, and cancer research. The protein's involvement in tumor growth, angiogenesis, and epithelial cell proliferation makes it a valuable tool for studying cancer biology and developing potential anticancer strategies.

To reconstitute and prepare Recombinant Human TGF-α protein for cell culture experiments, follow these steps for optimal protein stability and biological activity:

1. Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.

2. Reconstitution:

   • Most protocols recommend reconstituting the protein in sterile water or sterile 10 mM acetic acid.
   • Typical reconstitution concentrations are 0.1–1.0 mg/mL. For example, dissolve 100 μg in 1 mL for 0.1 mg/mL, or in 100 μL for 1.0 mg/mL.
   • If using acetic acid, add the appropriate volume of sterile 10 mM acetic acid to achieve the desired concentration.

3. Mixing:

   • Gently pipette the solution down the sides of the vial to dissolve the protein. Do not vortex as this can denature the protein.
   • Allow several minutes for complete dissolution.

4. Aliquoting and Storage:

   • For short-term use (up to 1 week), store the reconstituted protein at 2–8 °C.
   • For long-term storage, aliquot the solution (to avoid repeated freeze-thaw cycles) and store at –20 °C to –80 °C.
   • To enhance stability during storage, especially for long-term, add a carrier protein such as 0.1% BSA or HSA to the aliquots.

5. Dilution for Cell Culture:

   • Before adding to cell cultures, dilute the reconstituted stock to the desired working concentration using cell culture medium or other suitable aqueous buffer.
   • If the stock was reconstituted in acetic acid, ensure the final acetic acid concentration in the culture medium is negligible to avoid cytotoxicity.

Additional Best Practices:

• Avoid repeated freeze-thaw cycles, as these can reduce protein activity.
• Always consult the product-specific datasheet for any unique instructions regarding your particular recombinant TGF-α preparation.

Summary Table:

These guidelines will help ensure maximum stability and bioactivity of recombinant human TGF-α for your cell culture experiments.