Recombinant Human FGF-8a

Recombinant Human FGF-8a is a valuable tool for research due to its well-documented roles in regulating key cellular processes and developmental pathways. Here are the main reasons to use Recombinant Human FGF-8a in your research applications:

1. Regulation of Embryogenesis and Development

FGF-8a is essential for embryonic development, including the formation of the midbrain/hindbrain, limbs, eyes, ears, and heart. It mediates epithelial-mesenchymal transitions and plays an organizing and inducing role during gastrulation. This makes it critical for studies on developmental biology and organogenesis.

2. Cell Proliferation, Differentiation, and Migration

FGF-8a is a potent mitogen that stimulates cell proliferation, differentiation, and migration. It activates Ras/MAPK signaling pathways, which are central to these processes. This property is useful for experiments involving stem cell expansion, tissue regeneration, and cancer research.

3. Neural Stem Cell and iPSC Differentiation

FGF-8a is widely used to differentiate induced pluripotent stem cells (iPSCs), embryonic stem cells (ESCs), and neural stem cells. It supports the generation of specific neural lineages and is frequently applied in protocols for modeling neurological diseases and developmental disorders.

4. High Purity and Biological Activity

Recombinant Human FGF-8a is typically available with high purity (>95–98%) and low endotoxin levels, ensuring reliable and reproducible results in sensitive cell-based assays. Its biological activity is validated using standardized proliferation assays (e.g., NR6R-3T3 or BALB/c 3T3 cells), with ED₅₀ values in the range of 0.1–2 μg/mL.

5. Relevance to Disease Models

FGF-8a has been implicated in tumor growth, angiogenesis, and certain cancers (e.g., prostate, breast, ovarian). Its expression is also linked to tissue repair and wound healing, making it relevant for studies on cancer biology, regenerative medicine, and tissue engineering.

6. Well-Characterized and Widely Cited

Recombinant Human FGF-8a is supported by extensive scientific literature and product validation data. Its use is documented in numerous peer-reviewed publications, ensuring that your research is built on a solid and credible foundation.

7. Versatile Applications

It is suitable for a wide range of applications, including:

• Bioassays and cell proliferation studies
• Stem cell differentiation protocols
• Organoid and 3D culture systems
• Disease modeling (e.g., neurological disorders, cancer)
• Drug discovery and screening

8. Species Compatibility

Recombinant Human FGF-8a is active in human, mouse, and other mammalian cell systems, making it broadly applicable across different experimental models.

In summary, Recombinant Human FGF-8a is a versatile, well-characterized growth factor that supports a wide array of research applications, particularly in developmental biology, stem cell research, and disease modeling. Its high purity, biological activity, and relevance to key cellular processes make it an essential reagent for advanced scientific studies.

Recombinant Human FGF-8a can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity, well-characterized, and compatible with your assay system.

Key considerations and supporting details:

• Purity and Characterization: Recombinant Human FGF-8a proteins are typically supplied at >95% or >98% purity, confirmed by SDS-PAGE, and are biologically active as demonstrated by cell proliferation assays. This level of purity is generally suitable for use as a standard in quantitative assays such as ELISA.

• Carrier-Free vs. Carrier-Added: For ELISA calibration, both carrier-free and carrier-added (e.g., BSA) forms are available. Carrier-added forms are often recommended for use as ELISA standards because they enhance protein stability and reduce adsorption losses, especially at low concentrations. However, if your assay is sensitive to BSA or other carriers, use the carrier-free version.

• Standard Curve Preparation: To use recombinant FGF-8a as a standard, reconstitute it according to the supplier’s instructions (commonly in sterile PBS or assay buffer), and prepare a serial dilution to generate a standard curve covering the expected concentration range in your samples. Ensure the diluent matches your sample matrix to minimize matrix effects.

• Assay Compatibility: Confirm that your ELISA kit or custom assay is specific for the FGF-8a isoform and does not exhibit significant cross-reactivity with other FGF family members or isoforms. Most commercial FGF8 ELISA kits are designed for total FGF8, but some may distinguish between isoforms.

• Validation: For quantitative accuracy, validate the standard curve by assessing linearity, recovery, and parallelism with your sample matrix. This ensures that the recombinant standard behaves similarly to endogenous FGF-8a in your assay conditions.

• Documentation: Record the lot number, concentration, and reconstitution details of the recombinant standard for reproducibility and traceability.

Summary Table: Key Requirements for Using Recombinant FGF-8a as an ELISA Standard

Conclusion:
You can use recombinant human FGF-8a as a standard for ELISA quantification, provided it is validated for your specific assay and sample type, and you follow best practices for standard preparation and assay validation.

Recombinant Human FGF-8a has been validated for several key applications in published research, primarily in the context of cell culture, stem cell biology, and developmental studies. The main applications supported by published data include:

1. Cell Proliferation Assays:
   Recombinant Human FGF-8a has been shown to stimulate cell proliferation in various cell lines, such as the NR6R-3T3 mouse fibroblast cell line, with an ED50 typically ranging from 0.25–1.5 μg/mL in the presence of heparin. This application is widely used for bioassays and functional validation of FGF-8a activity.

2. Stem Cell Differentiation:
   FGF-8a is critical for the differentiation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells into specific neural lineages, particularly ventral midbrain dopaminergic (VM DA) neurons. Studies have demonstrated that early exposure to FGF-8a (rather than FGF-8b) is required for robust differentiation of FOXA2+ floor plate-like neural progenitors, which are essential for Parkinson’s disease modeling and cell-based therapies.

3. Pluripotency Maintenance:
   FGF-8a is used in protocols for maintaining pluripotency in human pluripotent stem cells, supporting their undifferentiated state during culture.

4. Endoderm Induction and Characterization:
   FGF-8a is applied in protocols for endoderm induction and characterization in hPSC-derived endoderm cell research, highlighting its role in early developmental processes.

5. Neural Development and Organoid Models:
   FGF-8a has been used in neural cultures and organoid models to study neuronal morphogenesis and developmental defects, such as in models of Leigh syndrome and Parkinson’s disease.

6. Cancer and Tumor Biology:
   FGF-8a has been implicated in tumor growth and angiogenesis, with studies showing its expression in various human tumors (e.g., prostate, breast, ovarian). It is used in research to investigate the role of FGF signaling in tumorigenesis and cancer progression.

7. Developmental Biology:
   FGF-8a is essential for various developmental processes, including gastrulation, somitogenesis, morphogenesis, and limb development, as demonstrated in both in vitro and in vivo models.

These applications are supported by primary research articles and product validation data from multiple sources, confirming the utility of Recombinant Human FGF-8a in a wide range of scientific studies.

To reconstitute and prepare Recombinant Human FGF-8a protein for cell culture experiments, dissolve the lyophilized protein in sterile water or buffer to a concentration of at least 100 μg/mL, incubate at room temperature for 20 minutes, and avoid vortexing to preserve biological activity. After reconstitution, further dilute in cell culture medium or buffer containing a carrier protein such as 0.1% BSA or 10% FBS to stabilize the protein for experimental use.

Step-by-step protocol:

• Before opening the vial: Centrifuge at 3000 rpm for 5 minutes to collect all protein at the bottom.
• Reconstitution:
  • Add sterile H₂O or PBS to achieve a final concentration of ≥100 μg/mL (some protocols recommend up to 500 μg/mL in PBS for stock solutions).
  • Gently mix by pipetting; do not vortex, as vigorous agitation may reduce activity.
  • Incubate at room temperature for at least 20 minutes to ensure complete dissolution.
• Aliquoting and storage:
  • Aliquot the stock solution to minimize freeze/thaw cycles.
  • Store aliquots at −20°C or −80°C for long-term use (up to 3–6 months); short-term storage (up to 1 week) can be at 2–8°C.
• Working solution preparation:
  • Dilute the stock solution in cell culture medium or buffer containing a carrier protein (e.g., 0.1% BSA, 10% FBS, or 5% HSA) to the desired working concentration for your assay.
  • Typical effective concentrations for cell proliferation assays range from 0.1–3 μg/mL, depending on cell type and experimental design.
• Additional notes:
  • Avoid repeated freeze/thaw cycles to maintain protein integrity.
  • Use reconstituted protein within one month for best results.
  • Confirm protein activity by a cell proliferation assay if needed.

Summary of key points:

• Reconstitute in sterile H₂O or PBS, ≥100 μg/mL.
• Incubate at room temperature for 20 minutes.
• Do not vortex.
• Dilute in buffer with carrier protein for cell culture.
• Store aliquots at −20°C or −80°C; avoid freeze/thaw cycles.

These steps ensure optimal solubility, stability, and biological activity of recombinant FGF-8a for cell culture experiments.