Recombinant Human TFF2

Recombinant Human TFF2 is used in research applications primarily for its roles in mucosal protection, epithelial repair, anti-inflammatory activity, and modulation of immune responses, making it valuable for studies on gastrointestinal, respiratory, and other mucosal tissue injuries and diseases.

Key scientific reasons to use recombinant human TFF2 include:

• Mucosal Healing and Epithelial Repair: TFF2 accelerates repair in models of gastric and mucosal injury, demonstrating cytoprotective effects and promoting epithelial regeneration and wound healing.
• Anti-inflammatory Properties: TFF2 can attenuate inflammatory responses, which is beneficial in models of respiratory and gastrointestinal inflammation.
• Immune Modulation: TFF2 acts as a chemoattractant for human monocytes, indicating a role in immune cell recruitment and mucosal immune regulation.
• Gastrointestinal Function: TFF2 inhibits gastrointestinal motility and gastric acid secretion, and may stabilize gastric mucus, supporting studies on gut physiology and pathology.
• Respiratory Applications: Recombinant TFF2, especially when fused with interferons, has shown promise in reducing morbidity and mortality in viral respiratory infections (e.g., influenza, COVID-19) by repairing pulmonary function and regulating immune responses.
• Safety and Delivery: Clinical studies indicate that recombinant TFF2 (and TFF2-IFN fusion proteins) are well-tolerated, with prolonged retention in mucosal tissues and minimal systemic exposure when administered by inhalation, making them suitable for localized therapeutic research.

Typical research applications include:

• In vitro and in vivo models of mucosal injury and repair.
• Studies of inflammatory bowel disease, peptic ulcers, and other gastrointestinal disorders.
• Investigations into respiratory tract injury and viral infections.
• Assays for immune cell chemotaxis and cytokine modulation.
• ELISA, Western blot, immunoprecipitation, and functional cell-based assays.

Summary Table: Scientific Functions of Recombinant Human TFF2

Using recombinant human TFF2 enables precise mechanistic studies and therapeutic modeling in mucosal biology, inflammation, and tissue repair, supporting translational research in both gastrointestinal and respiratory disease contexts.

Recombinant Human TFF2 can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined. This is a common practice in both commercial ELISA kits and custom assay development.

Supporting details:

• Commercial ELISA kits for TFF2 routinely use recombinant human TFF2 as the standard for generating calibration curves, enabling quantitative measurement of TFF2 in biological samples. The standard is typically a recombinant protein, often produced in E. coli or mammalian cells, and its concentration is carefully validated for use in the assay.
• Recombinant TFF2 protein standards are specifically marketed for use in ELISA calibration, and protocols indicate their suitability for this purpose. For example, recombinant human TFF2 expressed in HEK293 cells or E. coli is used as a standard in both research and commercial settings.
• Best practices:
  • Ensure the recombinant TFF2 is of known and verified concentration (preferably mass-calibrated or value-assigned using a reference method).
  • The standard curve should be freshly prepared for each assay to ensure accuracy.
  • If using a recombinant protein not supplied with your ELISA kit, it is advisable to validate its performance in your specific assay system, as differences in folding, post-translational modifications, or tags may affect antibody recognition and quantification.
  • Some sources recommend using recombinant proteins with carrier proteins (e.g., BSA) for stability, especially if the standard is to be stored or used over multiple assays.

Limitations and considerations:

• If your ELISA kit is designed for a specific form or tag of TFF2, ensure your recombinant standard matches this as closely as possible to avoid discrepancies in quantification.
• Always check the documentation of your ELISA kit for any specific requirements or recommendations regarding the standard protein.

In summary:
You can use recombinant human TFF2 as a standard for ELISA quantification, provided it is well-characterized, pure, and its concentration is accurately known. This approach is standard in both commercial and custom ELISA protocols.

Recombinant Human TFF2 has been validated for several scientific applications in published research, primarily in the fields of gastrointestinal biology, mucosal repair, immunology, and respiratory disease models.

Validated Applications:

• Functional Assays: Used to assess biological activity, such as cytoprotection, chemotaxis, and modulation of epithelial cell behavior.
• Blocking Assays: Applied to study inhibition of specific biological pathways or interactions.
• ELISA (Enzyme-Linked Immunosorbent Assay): Utilized as a standard or antigen for quantifying TFF2 levels in biological samples.
• Western Blot: Used as a positive control or for antibody validation in protein detection.
• SDS-PAGE: Employed for purity assessment and molecular weight determination.
• HPLC (High-Performance Liquid Chromatography): Used for protein characterization and purity analysis.

Published Research Applications:

• Gastrointestinal Mucosal Repair: Recombinant human TFF2 has been shown to accelerate healing and provide cytoprotection in models of gastric injury, including burn-induced gastric lesions in mice.
• Epithelial Regeneration and Wound Healing: TFF2 plays a role in promoting epithelial cell migration and repair, particularly in the gastrointestinal tract.
• Respiratory Disease Models: Recombinant TFF2, especially in fusion with interferon (TFF2-IFN), has been investigated for its therapeutic potential in respiratory viral infections (e.g., COVID-19, influenza), showing anti-inflammatory effects, mucosal repair, and reduced morbidity in animal models and early-phase clinical trials.
• Immunomodulation: Studies have explored TFF2's ability to regulate immune responses, including attenuation of inflammation and modulation of cytokine profiles.
• Oncology and Sepsis Research: TFF2 has been studied in the context of cancer biology and systemic inflammatory responses, though these applications are less extensively validated.

Key Biological Activities:

• Cytoprotection: Protects epithelial cells from injury and stress.
• Inhibition of Gastrointestinal Motility and Acid Secretion: May stabilize gastric mucus and interact with glycoproteins.
• Chemotaxis: Induces migration of specific cell types, such as MCF-7 cells, in bioassays.

Summary Table:

Note: While recombinant human TFF2 is widely used in research, its applications are primarily for preclinical and mechanistic studies, not for diagnostic or therapeutic procedures.

To reconstitute and prepare Recombinant Human TFF2 protein for cell culture experiments, dissolve the lyophilized protein in sterile, high-purity water (18 MΩ-cm H₂O) at a concentration of at least 100 µg/mL. This stock can then be further diluted in appropriate aqueous buffers or cell culture media as needed for your experiments.

Detailed protocol and best practices:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Add sterile water (or 0.1% BSA in water, if additional stability is needed) to achieve a concentration of at least 100 µg/mL. For example, add 100 µL water to 100 µg lyophilized protein for a 1 mg/mL solution.
• Gently mix by swirling or inverting the vial. Avoid vigorous shaking or vortexing to prevent foaming and protein denaturation.
• Allow the protein to fully dissolve at room temperature for 15–30 minutes with gentle agitation.
• If required, further dilute the reconstituted protein in cell culture medium or buffer to the desired working concentration. For cell-based assays, concentrations in the range of 1–10 ng/mL are commonly used for TFF2 bioactivity.
• For long-term storage, aliquot the reconstituted protein and store at –20°C or below. For short-term use (up to one week), store at 4°C. To enhance stability during storage, especially for long-term, add a carrier protein such as 0.1% BSA or HSA.
• Avoid repeated freeze-thaw cycles by aliquoting into single-use volumes.

Additional notes:

• The protein is typically supplied in a PBS-based lyophilization buffer, but always check the product datasheet for specific formulation details.
• If using for sensitive cell culture applications, ensure the final solution is sterile-filtered if necessary.
• Confirm the absence of endotoxin if required for your application (TFF2 preparations for cell culture are typically <1 EU/µg).

Summary Table: TFF2 Reconstitution for Cell Culture

Always consult the specific product datasheet for any unique instructions or formulation details.