Recombinant Human FGF-16

Recombinant Human FGF-16 is a valuable tool for a wide range of research applications due to its unique biological properties and multifaceted roles in cellular regulation. Here are several compelling reasons to consider using Recombinant Human FGF-16 in your research:

1. Versatile Receptor Binding and Cellular Effects

Recombinant Human FGF-16 binds to multiple fibroblast growth factor receptors (FGFRs), including FGFR1, FGFR2, and FGFR3. This broad receptor specificity allows it to influence a variety of cell types, making it highly versatile for studies involving cell proliferation, differentiation, and tissue regeneration.

2. Stem Cell Research

FGF-16 is widely used in stem cell research for its ability to:

• Promote the proliferation and differentiation of embryonic stem cells.
• Induce mesenchymal stem cells to differentiate into bone and cartilage cells, supporting studies in tissue engineering and regenerative medicine.

3. Cardiac Development and Repair

FGF-16 plays a critical role in embryonic heart development and adult cardiac homeostasis. It has been shown to:

• Mitigate pathological cardiac remodeling, such as fibrosis and hypertrophy.
• Exert cardioprotective effects by competitively inhibiting FGF2-induced signaling pathways, which are involved in adverse cardiac remodeling.

4. Metabolic Regulation

Emerging evidence highlights FGF-16's role in metabolic regulation:

• Overexpression of FGF-16 in rodent models leads to improved glucose tolerance and reduced insulin resistance.
• It influences lipid metabolism and can result in reduced fat mass and improved overall metabolic health.

5. Tissue Engineering and Regeneration

The mitogenic and differentiation-inducing properties of FGF-16 make it a valuable tool for:

• Enhancing the growth and differentiation of cells in engineered tissues.
• Promoting the regeneration of bone, cartilage, and nerve tissues.

6. Cancer Research

Aberrant expression of FGF-16 has been associated with various cancers. Recombinant FGF-16 can be used to:

• Study the mechanisms of cancer cell proliferation and invasion.
• Investigate its potential as a therapeutic target for cancer therapy, as it has been shown to inhibit cancer cell growth and induce cell death.

7. Wound Healing

FGF-16 promotes the proliferation and migration of skin cells, leading to faster wound closure and improved tissue regeneration. This makes it a potential therapeutic agent for:

• Treating chronic wounds.
• Promoting tissue repair in various experimental models.

8. Diagnostic and Biomarker Applications

Due to its ability to bind multiple FGFRs, Recombinant Human FGF-16 can be used as an antigen in diagnostic assays for various diseases. It has been shown to be a potential biomarker for certain cancers and can be utilized in immunoassays to detect FGF-16 in patient samples.

9. High Evolutionary Conservation

FGF-16 exhibits high sequence and structural conservation across vertebrates, indicating its essential biological functions. This conservation makes it a reliable and relevant protein for comparative studies across different species.

10. Research Use Only

Recombinant Human FGF-16 is available for research use only and is not approved for clinical diagnosis or human use, ensuring that it meets the regulatory requirements for laboratory research.

In summary, Recombinant Human FGF-16 is a powerful and versatile protein that can significantly enhance your research in areas such as stem cell biology, tissue engineering, cardiac development, metabolic regulation, cancer, and wound healing. Its broad receptor binding, well-documented biological effects, and high evolutionary conservation make it an excellent choice for a wide range of experimental applications.

Yes, recombinant human FGF-16 can be used as a standard for quantification and calibration in ELISA assays. This is a well-established practice in fibroblast growth factor research.

Suitability for ELISA Standards

Recombinant FGF-16 is specifically formulated for use as an ELISA standard. The protein is typically supplied in lyophilized form and requires reconstitution according to manufacturer specifications. For ELISA applications, recombinant FGF-16 with carrier protein (such as bovine serum albumin) is recommended, as the carrier protein enhances stability, extends shelf-life, and allows storage at more dilute concentrations.

Standard Curve Preparation

When preparing your standard curve, follow these guidelines:

Reconstitution and Concentration Range: Recombinant FGF-16 should be reconstituted in sterile phosphate-buffered saline (PBS) containing at least 0.1% human or bovine serum albumin. Standard curves typically range from 0 to 1000 pg/mL, though higher ranges up to 3000 pg/mL can be used if your target protein concentration is predicted to be extremely high.

Calibration Accuracy: The recombinant protein will generate linear calibration curves with high reproducibility. ELISA kits calibrated with recombinant standards demonstrate excellent linearity, with R² values typically exceeding 0.999 across the measurement range.

Storage and Handling

Store the reconstituted recombinant FGF-16 at the temperature recommended by the manufacturer, using a manual defrost freezer and avoiding repeated freeze-thaw cycles. This preservation of protein integrity is critical for maintaining the accuracy of your standard curve throughout your experiments.

The recombinant protein will reliably quantitate both natural and recombinant FGF-16 in your samples, making it suitable for accurate calibration of your ELISA assays.

Research Applications of Recombinant Human FGF-16

Recombinant human FGF-16 has been validated for several important research applications across multiple biological domains:

Cardiac Development and Homeostasis

FGF-16 is critically involved in embryonic heart development and adult cardiac homeostasis. Research demonstrates that FGF-16 significantly mitigates pathological cardiac remodeling, including fibrosis and hypertrophy, through competitive inhibition of FGF2-induced transforming growth factor-β1 signaling via FGF receptor 1c. The protein acts primarily through local paracrine signaling to mediate protective roles in the adult heart under stress or pathological conditions.

Cell Proliferation and Differentiation Studies

The biological activity of recombinant human FGF-16 has been validated through its ability to stimulate thymidine incorporation in quiescent fibroblasts, making it suitable for cell proliferation assays. FGF-16 plays an imperative role in the regulation of embryonic development, cell proliferation and cell differentiation, and is required for normal cardiomyocyte proliferation.

Bioassay Applications

FGF-16 has been validated for use in bioassay applications. The protein functions on cells of mesodermal and neuroectodermal origin to regulate diverse physiologic functions including angiogenesis, cell growth, pattern formation, embryonic development, metabolic regulation, cell migration, neurotrophic effects, and tissue repair.

Central Nervous System Development

FGF-16 plays a role in the development of the central nervous system, indicating its utility in neurodevelopmental research applications.

Metabolic Regulation Research

Emerging evidence highlights FGF-16's significant contribution to metabolic regulation, suggesting applications in studying metabolic disorders and related pathways.

Reconstitution Protocol

Pre-reconstitution preparation is critical for successful protein recovery. Before opening the lyophilized vial, centrifuge it at 3000 rpm for 5 minutes to consolidate any protein powder that may have adhered to the cap or vial walls. This ensures maximum protein recovery when you begin reconstitution.

Reconstitution buffer selection depends on your specific product formulation. Most FGF-16 preparations recommend reconstituting in sterile PBS (phosphate-buffered saline) at a concentration of 10 μg/mL, with the PBS containing at least 0.1% human or bovine serum albumin (BSA) as a carrier protein. Some formulations may specify alternative buffers such as sterile distilled water or Tris buffer (5 mM, pH 7.6). Always consult your product's Certificate of Analysis or data sheet for the exact reconstitution buffer recommended for your specific preparation.

Reconstitution procedure should be performed gently to preserve protein bioactivity. Add the appropriate volume of reconstitution buffer to the vial and allow the protein to dissolve at room temperature for at least 20 minutes. Do not vortex the solution, as vigorous shaking can impair biological activity. Instead, gently mix the vial after reconstitution. The protein may initially appear as a film at the bottom of the vial, which is normal.

Storage and Stability

Lyophilized protein storage is stable for 6-12 months when maintained at -20°C to -80°C in a desiccated state.

Reconstituted protein storage requires careful temperature management:

• Short-term storage (2-7 days): Store at 2-8°C under sterile conditions
• Extended storage (3-6 months): Store aliquots at -20°C or -80°C
• Long-term stability: Up to 3 months at -20°C or below

Critical handling consideration: Avoid repeated freeze-thaw cycles, as these can significantly reduce protein activity and stability. Use a manual defrost freezer and prepare aliquots in appropriate volumes to minimize the need for repeated thawing.

Optimization for Cell Culture

To enhance protein stability during cell culture experiments, further dilute your reconstituted FGF-16 in buffers containing carrier proteins or stabilizers such as 0.1% BSA, 10% fetal bovine serum (FBS), 5% human serum albumin (HSA), or 5% trehalose solution. This is particularly important since FGF-16 can lose activity in purely aqueous solutions.

The specific bioactivity of recombinant human FGF-16 is typically greater than 3 × 10⁴ IU/mg, with an ED₅₀ for inducing 3T3 cell proliferation of less than 31 ng/mL. This high specific activity means you can achieve effective results with relatively low protein concentrations in your cell culture experiments.