Recombinant Human FGF-Basic (146 aa)

Recombinant Human FGF-Basic (146 aa) is widely used in research due to its potent mitogenic and angiogenic activities, making it essential for studies involving cell proliferation, tissue regeneration, and developmental biology.

FGF-Basic (also known as FGF-2 or bFGF) is a member of the fibroblast growth factor family and is involved in several key biological processes:

• Cell Proliferation: Stimulates the growth of cells from mesodermal, neuroectodermal, ectodermal, and endodermal origins, making it valuable for expanding stem cells, fibroblasts, endothelial cells, and other primary cultures.
• Angiogenesis: Promotes the formation of new blood vessels, which is critical for wound healing, tissue engineering, and cancer research.
• Embryonic Development: Plays a central role in morphogenesis and differentiation during embryogenesis, supporting studies in developmental biology and organoid models.
• Wound Healing and Tissue Repair: Enhances regeneration and repair of various tissues, including skin, muscle, cartilage, and nervous tissue.
• Neuronal Function: Induces neuronal differentiation, survival, and regeneration, making it useful for neurobiology and disease modeling.
• Pathological Processes: Involved in tumor growth and inflammation, relevant for cancer biology and immunology research.

Technical Advantages:

• The 146 amino acid recombinant form is the full-length human FGF-2 isoform, ensuring broad biological activity and compatibility with human cell systems.
• Recombinant production allows for high purity (>95%), low endotoxin levels, and batch-to-batch consistency, which are critical for reproducible experimental results.
• Animal-free formulations are available, minimizing experimental variability and risk of contamination from animal-derived components.

Common Applications:

• Cell culture supplementation for stem cell maintenance and expansion.
• Bioassays for cell proliferation and differentiation.
• Organoid and tissue engineering protocols.
• Angiogenesis and wound healing models.
• Disease modeling, including neurodegeneration and cancer.

Best Practices:

• Avoid repeated freeze/thaw cycles to preserve protein activity.
• Select carrier-free or animal-free formulations if downstream applications are sensitive to additives or animal-derived contaminants.

In summary, Recombinant Human FGF-Basic (146 aa) is a versatile and essential growth factor for research in cell biology, regenerative medicine, developmental biology, and disease modeling due to its broad spectrum of biological activities and technical reliability.

Yes, recombinant human FGF-Basic (146 aa) can be used as a standard for quantification and calibration in ELISA assays. This is one of the primary recommended applications for this protein formulation.

Formulation Selection for ELISA

When using recombinant FGF-Basic for ELISA standardization, you should select the formulation with BSA (Bovine Serum Albumin) as a carrier protein. The carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at more dilute concentrations—all critical factors for maintaining standard integrity across multiple assay runs.

Alternatively, if you require a carrier-free formulation, this option is available for applications where the presence of BSA could interfere with your specific assay design.

Protein Specifications

The 146 amino acid variant has the following characteristics relevant to ELISA applications:

• Molecular weight: Approximately 16.5 kDa (predicted), migrating at ~17 kDa on SDS-PAGE
• Purity: Greater than 95% by SDS-PAGE analysis
• Endotoxin level: Less than 0.1 EU/µg, determined by LAL method
• Formulation: Typically supplied as a 0.22 µm filtered protein solution in pH 7.5 Tris buffer with 150 mM NaCl

Recommended Dilution Range

For ELISA applications, typical working dilutions range from 0.2 to 0.4 ng/well, depending on your specific assay optimization requirements.

Important Consideration

Note that ELISA standard formulations are specifically validated for quantification purposes and should not be used interchangeably with bioassay-grade recombinant proteins, as ELISA standards are not tested for cell proliferation or functional bioassay applications.

Research Applications of Recombinant Human FGF-Basic (146 aa)

Recombinant human FGF-basic (146 aa) has been validated across a diverse range of research applications in published literature, reflecting its broad biological activity and utility in multiple experimental contexts.

Stem Cell Biology and Differentiation

FGF-basic (146 aa) has demonstrated significant utility in stem cell research. The protein supports neural stem cell expansion, with documented use at 20 ng/mL concentrations achieving 100-fold expansion of rat and mouse cortical stem cells over multiple passages. In pluripotent stem cell applications, the protein has been employed for directed differentiation into specialized cell types, including lung-specific mesenchyme, Schwann cells, and germ-like cells. The protein also supports the development of functional human pluripotent stem cell-derived Kupffer cells in hepatic organoid systems.

Cell Proliferation and Bioassays

The primary validated application involves cell proliferation assays, where recombinant human FGF-basic (146 aa) induces dose-dependent proliferation of NIH/3T3 mouse fibroblasts. This bioassay application has been extensively utilized across numerous published studies examining cellular responses to growth factor signaling. The protein stimulates proliferation of cells derived from mesodermal, neuroectodermal, ectodermal, and endodermal origins.

Organoid Development and Tissue Engineering

FGF-basic (146 aa) has been validated for use in regenerative human liver organoid (HLO) development for hepatotoxicity assays. The protein supports organoid formation and maturation in perfusion-based culture systems, enabling functional tissue modeling.

Neurobiological Research

The protein has been applied in studies examining neuronal differentiation, survival, and regeneration. Published research has utilized FGF-basic in investigations of neural development, neurodegeneration, and disease modeling in neurological conditions.

Disease Modeling Applications

FGF-basic (146 aa) has been validated in multiple disease modeling contexts, including studies of Parkinson's disease, glioblastoma, synovial sarcoma, skeletal dysplasia, and Hirschsprung's disease. The protein supports the generation and maintenance of disease-relevant cell populations for mechanistic studies and therapeutic screening.

Tissue Culture and Maintenance

The protein is used to supplement tissue culture media for supporting growth of specialized cell populations, including human and canine chondrocyte-like cells.

To reconstitute and prepare Recombinant Human FGF-Basic (146 aa) protein for cell culture experiments, dissolve the lyophilized protein at 100–250 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA or HSA). This carrier protein is essential to stabilize FGF-Basic and prevent adsorption to surfaces and loss of activity.

Step-by-step protocol:

• Briefly centrifuge the vial before opening to collect all lyophilized material at the bottom.
• Add sterile PBS (pH 7.2–7.4) containing ≥0.1% BSA or HSA to achieve the desired concentration (typically 100–250 μg/mL).
• Gently mix by swirling or tapping the vial; avoid vigorous pipetting or vortexing to prevent protein denaturation.
• Allow the protein to dissolve completely (usually 5–10 minutes at room temperature).
• If necessary, further dilute the stock solution in cell culture medium or buffer containing carrier protein (≥0.1% BSA/HSA) to reach your working concentration.
• Aliquot the reconstituted solution to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at –20°C or colder for long-term storage, or at 2–8°C for short-term use (up to one week).

Additional notes:

• If the product is carrier-free, you must add BSA or HSA during reconstitution to maintain stability.
• Avoid using purely aqueous solutions without carrier protein, as FGF-Basic is prone to aggregation and loss of bioactivity.
• For bioassays, typical working concentrations range from 10–100 ng/mL depending on cell type and application.
• Always use sterile technique to prevent contamination.

Summary Table:

This protocol ensures optimal stability and bioactivity of FGF-Basic for cell culture experiments.