Trefoil factor 1, also known as TFF1 is a small cysteine-rich secreted protein that is frequently expressed in breast tumors.1 It is also normally expressed in the stomach, and is found ectopically in gastrointestinal inflammatory disorders and in various carcinomas. It is involved in stomach ontogenesis and in the maintenance of the integrity of the mucosa, and may represent a pharmacological tool for prevention and healing of gastrointestinal ulcerations. TFF1 is a pleiotropic factor involved in mucin polymerization, cell motility, cell proliferation and/or differentiation, and possibly in the nervous system.2
Protein Details
Purity
>98% by SDS Page and HPLC
Endotoxin Level
<1.0 EU/µg
Biological Activity
The ED50 as determined by a chemotaxis bioassay using human MCF-7 cells is less than 10µg/ml, corresponding to a specific activity of >100 IU/mg.
The Human TFF1 protein was lyophilized from a 0.2µm filtered concentrated solution in 20mM PB, pH 7.2 - 7.4 and 150mM NaCl.
Reconstitution
For reconstitution, it is advised to use sterile high-purity water (18MΩ-cm) with the lyophilized protein at a concentration of at least 100µg/ml. Subsequent dilutions into other aqueous solutions can be performed after this initial step.
Storage and Stability
The lyophilized protein should be stored desiccated at -20°C. The reconstituted protein can be stored for at least one week at 4°C. For long-term storage of the reconstituted protein, aliquot into working volumes and store at -20°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
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Recombinant Human TFF1 is widely used in research due to its critical roles in epithelial protection, regeneration, wound healing, and its involvement in cancer biology and cellular signaling pathways.
Key scientific reasons to use Recombinant Human TFF1:
Epithelial Regeneration and Wound Healing: TFF1 promotes epithelial cell motility and restitution, making it valuable for studies on mucosal repair, tissue engineering, and regenerative medicine. Its motogenic properties can be exploited to enhance healing in models of epithelial injury.
Mucosal Integrity: TFF1 is essential for maintaining the integrity of gastric and intestinal mucosa. It is expressed by goblet cells and interacts with mucins, contributing to the physical protection of mucosal surfaces.
Cellular Signaling: TFF1 modulates key signaling pathways such as ERK/MAPK and PI3K/Akt, which are crucial for cell division, differentiation, apoptosis, and survival. This makes it a useful tool for dissecting molecular mechanisms underlying these processes.
Cancer Research: TFF1 exhibits context-dependent roles in cancer:
Tumor Suppressor: In gastric and lung carcinoma, TFF1 suppresses proliferation, migration, and invasion, and induces apoptosis, suggesting its utility in studies of tumor suppression and cancer therapy.
Oncogenic Functions: In breast cancer, TFF1 enhances cell proliferation, migration, invasion, and tumor growth, making it relevant for investigating mechanisms of cancer progression and potential therapeutic targets.
Disease Biomarker: Altered TFF1 expression is associated with various diseases, including gastric cancer (down-regulation), breast and prostate cancers (up-regulation), and kidney stone formation (reduced urinary excretion). Recombinant TFF1 can be used to develop diagnostic assays or study disease mechanisms.
Protein-Protein Interactions: TFF1 forms complexes with other proteins (e.g., mucins, Gastrokine 2), allowing investigation of mucosal biology and protein interaction networks.
Experimental Versatility: Recombinant TFF1 enables controlled in vitro and in vivo studies, facilitating reproducible experiments in cell culture, animal models, and biochemical assays.
Typical research applications include:
Studying epithelial restitution and wound healing mechanisms.
Investigating mucosal barrier function and integrity.
Elucidating signaling pathways in cell proliferation, differentiation, and apoptosis.
Exploring TFF1’s dual roles in cancer biology.
Developing diagnostic and prognostic assays for mucosal and oncological diseases.
Engineering bio-scaffolds for regenerative medicine using TFF1’s motogenic properties.
In summary, recombinant human TFF1 is a versatile tool for research in epithelial biology, cancer, regenerative medicine, and disease biomarker development, owing to its multifaceted biological functions and relevance in health and disease.
Yes, you can use recombinant human TFF1 as a standard for quantification or calibration in your ELISA assays, provided it is of high purity and its concentration is accurately determined. This is a common and accepted practice in quantitative ELISA protocols for TFF1 and other proteins.
Key considerations and supporting details:
Recombinant TFF1 is widely used as an ELISA standard: Many commercial TFF1 ELISA kits use recombinant human TFF1 as the calibrator or standard for generating the standard curve. The standard is typically a purified recombinant protein, sometimes with a tag (e.g., His-tag), and is supplied with a known concentration.
Purity and quantification: For accurate calibration, the recombinant TFF1 should be highly purified and its concentration precisely quantified, typically by absorbance at 280 nm or another validated method. Impurities or inaccurate concentration measurements can lead to errors in quantification.
Expression system and post-translational modifications: Recombinant TFF1 can be expressed in various systems (e.g., E. coli, HEK293 cells). If your ELISA antibodies are sensitive to specific conformations or post-translational modifications, ensure your recombinant standard matches the native protein as closely as possible.
Carrier proteins: Some recombinant standards are supplied with carrier proteins (e.g., BSA) to enhance stability, while others are carrier-free. Choose the format that best matches your assay requirements and avoid carriers if they may interfere with detection.
Standard curve preparation: Prepare a serial dilution of the recombinant TFF1 standard to cover the expected concentration range in your samples (commonly 0–1000 pg/mL, but this may vary by kit). Always follow best practices for pipetting and dilution to ensure accuracy.
Validation: If you are developing a custom ELISA or using a non-kit standard, validate that your recombinant TFF1 is recognized equivalently to native TFF1 by your assay antibodies. This ensures the standard curve accurately reflects sample concentrations.
Summary Table: Use of Recombinant TFF1 as ELISA Standard
Requirement
Details/Best Practice
Protein source
Recombinant human TFF1 (high purity, accurately quantified)
Expression system
E. coli, HEK293, or other (match to assay if possible)
Carrier protein
With or without BSA (choose based on assay compatibility)
Standard curve range
Typically 0–1000 pg/mL (adjust as needed)
Validation
Confirm recognition by assay antibodies; compare to native TFF1 if possible
Preparation
Serial dilutions, fresh pipette tips, use within recommended time frame
In summary: Recombinant human TFF1 is suitable and commonly used as a standard for ELISA quantification, provided it is well-characterized and compatible with your assay system.
Recombinant Human TFF1 has been validated for several applications in published research, primarily in the fields of cancer biology, gastrointestinal protection, biomarker development, and nephrology.
Key validated applications include:
Cellular Functional Assays (in vitro):
Used to study cell proliferation, migration, invasion, and survival in mammary carcinoma and colorectal adenocarcinoma cell lines. Recombinant TFF1 has been shown to enhance oncogenicity, promote anchorage-independent growth, and increase cell motility in vitro.
Applied in wound healing and epithelial regeneration assays to assess its role in promoting epithelial cell motility and repair.
Animal Models (in vivo):
Used in xenograft models to demonstrate its effect on tumor growth and regression when neutralized by antibodies.
Employed in rat models of intracerebral hemorrhage to study its protective effects on cerebral edema and gastric mucosal injury, implicating the EGFR/Src/FAK pathway.
Biochemical and Biophysical Assays:
Utilized in crystal growth inhibition assays to show that TFF1 inhibits calcium oxalate crystal formation, relevant to nephrolithiasis research.
Used in studies of protein-protein interactions, such as binding to mucins (e.g., MUC5AC) and Gastrokine 2, and copper-mediated homodimerization.
Immunoassays and Biomarker Studies:
ELISA and immunohistochemistry: Recombinant TFF1 is used as a standard or control in ELISA assays for quantifying TFF1 in serum and tissue samples, particularly in studies of breast and gastric cancer.
Serum biomarker validation: Elevated serum TFF1 levels have been correlated with breast cancer and H. pylori infection, and its measurement is used for diagnostic and prognostic purposes.
Gastrointestinal Research:
Investigated for its therapeutic potential in treating and preventing gastrointestinal disorders associated with mucosal damage, due to its role in stabilizing the mucus layer and promoting mucosal healing.
Summary Table: Applications of Recombinant Human TFF1 in Published Research
ELISA, immunohistochemistry, serum biomarker validation for cancer and infection
Gastrointestinal Research
Mucosal protection, therapeutic studies for GI disorders
These applications are supported by multiple peer-reviewed studies and reviews, demonstrating the broad utility of recombinant human TFF1 in both basic and translational research.
Reconstitution Procedure
Recombinant Human TFF1 is typically supplied in lyophilized form and requires proper reconstitution before use in cell culture experiments. Begin by briefly centrifuging the vial to bring the contents to the bottom prior to opening. Allow 15-30 minutes for reconstitution at room temperature with gentle agitation, avoiding vigorous shaking that can cause foaming and protein denaturation.
The recommended reconstitution concentration ranges from 0.5-1.0 mg/mL using sterile deionized water or high-purity water (18 MΩ-cm), though some protocols recommend starting at concentrations not less than 100 µg/mL, which can then be further diluted to other aqueous solutions as needed. Alternatively, phosphate-buffered saline (PBS) can be used as a reconstitution medium at 500 μg/mL.
Storage and Stability Considerations
Lyophilized protein storage: Store the desiccated powder at temperatures below -18°C to -20°C. Although lyophilized TFF1 remains stable at room temperature for approximately 3 weeks, long-term storage at freezer temperatures is strongly recommended.
Reconstituted protein storage: Once reconstituted, store the protein at 4°C for short-term use. The reconstituted protein remains stable for at least one week under these conditions. For extended storage or to minimize degradation, prepare aliquots and store them at -80°C.
Critical handling: Avoid repeated freeze-thaw cycles, as these can compromise protein integrity and biological activity. When using the protein during experiments, remove only the amount needed from refrigeration and add it directly to culture medium rather than repeatedly accessing the stock solution.
Preparation for Cell Culture Applications
When preparing TFF1 for cell culture experiments, centrifuge cell culture supernatants at 2,000 × g for 10 minutes to remove debris before assaying. Dilute samples as necessary into appropriate sample diluent buffers. The protein can be directly added to culture medium at the desired working concentration based on your experimental design.
References & Citations
1. Westley BR et al. (2002) FASEB
2. Rio MC et al. (1998) Biochim Biophys Acta.1378: F61
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
