Recombinant Human TGF-β2 (Mammalian Derived)

Recombinant Human TGF-β2 (Mammalian Derived) is used in research applications because it is a biologically active, pleiotropic cytokine that regulates key cellular processes such as proliferation, differentiation, migration, apoptosis, and extracellular matrix production, making it essential for studies in developmental biology, tissue engineering, immunology, and disease modeling.

Key reasons to use mammalian-derived recombinant TGF-β2 include:

• Native Post-Translational Modifications: Mammalian expression systems provide post-translational modifications (e.g., glycosylation, proper folding, disulfide bond formation) that are critical for the full biological activity and receptor interactions of TGF-β2, closely mimicking the endogenous human protein.
• Functional Relevance: TGF-β2 is involved in regulating chondrocyte proliferation, differentiation, and migration, making it vital for cartilage biology, stem cell research, and regenerative medicine. It also plays roles in wound healing, immune modulation, fibrosis, and cancer progression.
• Cell-Type Specific Effects: TGF-β2 can inhibit epithelial cell proliferation, induce mesenchymal cell proliferation, and modulate immune responses, allowing precise control in cell culture and tissue models.
• Disease Modeling: It is used to study mechanisms underlying diseases such as osteoarthritis, rheumatoid arthritis, fibrosis, glaucoma, and cancer, and to test therapeutic interventions targeting TGF-β signaling.
• Bioassay and Functional Studies: Recombinant TGF-β2 is commonly used in bioassays to assess cell signaling, differentiation, and migration, and to validate pathway-specific effects in vitro.

Applications include:

• Inducing or inhibiting epithelial-mesenchymal transition (EMT) in cancer and fibrosis models.
• Promoting chondrogenesis and cartilage repair in tissue engineering.
• Modulating immune cell function in immunology studies.
• Investigating wound healing and tissue remodeling mechanisms.
• Studying angiogenesis and vascular biology.

Using a mammalian-derived recombinant protein ensures higher fidelity to native human TGF-β2, reducing variability and artifacts that may arise from non-mammalian expression systems, which is critical for reproducibility and translational relevance in biomedical research.

Yes, recombinant human TGF-β2 (mammalian derived) can generally be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity, properly quantified, and biologically active. This is a common practice in ELISA development and commercial kits, but several critical factors must be considered to ensure accurate and reliable results.

Key considerations and supporting details:

• Recombinant Protein as Standard: Most commercial TGF-β2 ELISA kits use recombinant human TGF-β2 as the standard for generating calibration curves. The standard is typically supplied as a lyophilized recombinant protein, which is reconstituted and serially diluted to create a standard curve for quantification.

• Source and Expression System: Mammalian-derived recombinant TGF-β2 is preferred for ELISA standards because it is more likely to have correct folding and post-translational modifications, closely resembling the native human protein. This improves antibody recognition and assay accuracy.

• Purity and Activity: The recombinant standard should be highly pure (typically >95%) and biologically active. Impurities or inactive protein can lead to inaccurate quantification.

• Quantification and Handling: The concentration of the recombinant standard must be accurately determined, usually by absorbance at 280 nm or amino acid analysis. Proper reconstitution (often in acidified buffer with carrier protein such as BSA) and storage conditions are essential to maintain stability and activity.

• Matrix Effects: When preparing the standard curve, dilute the recombinant protein in the same buffer or matrix as your samples to minimize matrix effects and ensure comparable binding kinetics.

• Validation: If you are developing your own ELISA or using a kit with a different standard, validate that your recombinant TGF-β2 standard produces a linear, reproducible standard curve and is recognized equivalently by the assay antibodies.

• Kit-Specific Recommendations: If you are using a commercial ELISA kit, always check the kit manual. Some kits are validated only with their supplied standard, and substituting another recombinant protein may affect accuracy unless equivalency is demonstrated.

Summary Table: Use of Recombinant TGF-β2 as ELISA Standard

In summary: You can use mammalian-derived recombinant human TGF-β2 as a standard for ELISA quantification, provided it is properly validated and handled according to best practices and any specific kit requirements.

Recombinant Human TGF-β2 (Mammalian Derived) has been validated in published research for several key applications, primarily involving bioassays, cell culture, and functional assays. These applications leverage its biological activity and relevance to human physiology and disease.

Validated Applications in Published Research:

• Bioassays:
  Used to assess cellular responses to TGF-β2, such as induction of epithelial-mesenchymal transition (EMT), modulation of cell signaling pathways, and measurement of downstream gene expression changes.
  Examples include:

  • Studying defective angiogenesis in microvessel-on-a-chip models.
  • Investigating TGF-β2-induced changes in trabecular meshwork cells and retinal pigment epithelium.

• Cell Culture:
  Applied as a supplement to cell culture media to study its effects on cell proliferation, differentiation, migration, and matrix production.
  Examples include:

  • Modulating cell spreading and focal adhesion formation.
  • Inducing myofibroblast transdifferentiation in conjunctival fibroblasts.
  • Supporting neuronal and vascular studies in hippocampal models.

• Functional Assays:
  Used to validate the biological activity of TGF-β2 in various cell-based systems, including HEK293-derived preparations for functional readouts.

Additional Research Contexts:

• Immunoassay Standard:
  Employed as a standard in immunoassays (e.g., ELISA) to quantify TGF-β2 levels in biological samples.

• Disease Modeling:
  Utilized in studies modeling fibrosis, cancer, wound healing, and immune regulation, reflecting its pleiotropic roles in health and disease.

• Protein Structure and Biochemistry:
  Used for structural studies, including recombinant production, purification, and crystallization to understand TGF-β2’s molecular properties.

Representative Published Studies:

• Angiogenesis and fibrosis: Microvessel-on-a-chip models for systemic sclerosis.
• Ophthalmology: Trabecular meshwork and retinal pigment epithelium studies.
• Cancer biology: Tumor microenvironment and resistance to kinase inhibitors.
• Neuroscience: Neuronal and vascular effects in hippocampal models.

Summary Table of Validated Applications

Key Notes:

• Most published research uses mammalian-derived TGF-β2 for its authentic glycosylation and biological activity, which are critical for functional studies.
• Applications are typically validated in human and mouse cell systems, with broad relevance to developmental biology, pathology, and therapeutic research.

If you need protocols or more specific details for a particular application, please specify the context or experimental system.

To reconstitute and prepare Recombinant Human TGF-β2 (Mammalian Derived) protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Reconstitution buffer: Most protocols recommend reconstituting in sterile, acidic buffer—commonly 4 mM HCl or 10 mM HCl—to a concentration of at least 20–100 μg/mL. Some protocols use sterile distilled water, but acidic conditions are preferred for stability and solubility.
• Carrier protein: If the protein is carrier-free, add at least 0.1% human or bovine serum albumin (BSA) to prevent adsorption to plastic and improve stability.
• Mixing: Gently swirl or tap the vial to dissolve. Do not vortex or shake vigorously, as this can denature the protein.
• Incubation: Allow the solution to sit at room temperature for 15–20 minutes to ensure complete dissolution.
• Aliquoting: After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage: Store reconstituted protein at 2–8°C for up to 1 week for short-term use, or at –20°C or below for long-term storage. Addition of carrier protein is recommended for long-term storage.
• Working dilutions: Prepare working concentrations for cell culture by diluting the stock solution in cell culture medium immediately before use. If using serum-free medium, ensure the presence of a carrier protein in the dilution buffer to maintain stability.

Example protocol:

1. Briefly centrifuge the vial.
2. Add sterile 4 mM HCl (with 0.1% BSA if carrier-free) to achieve a concentration of 100 μg/mL.
3. Gently swirl to dissolve; do not vortex.
4. Incubate at room temperature for 20 minutes.
5. Aliquot and store at –20°C for long-term use or 2–8°C for up to 1 week.
6. Dilute to desired working concentration in cell culture medium just before use.

Additional notes:

• If your application requires activation of latent TGF-β2, follow specific activation protocols (e.g., acidification and neutralization) as TGF-β2 is often secreted in a latent form.
• Always consult the product-specific Certificate of Analysis (CoA) or data sheet for any unique instructions.

These steps ensure maximum stability and biological activity of recombinant TGF-β2 for cell culture experiments.