Recombinant Human FGF-20

Recombinant Human FGF-20 is used in research primarily for its potent and selective effects on the survival, differentiation, and protection of midbrain dopaminergic neurons, making it highly relevant for studies in neurobiology, stem cell differentiation, and neurodegenerative disease modeling, especially Parkinson’s disease.

Key scientific applications and rationale include:

• Selective Promotion of Dopaminergic Neuron Differentiation: FGF-20 specifically enhances the differentiation of human embryonic stem cells (hESCs) into dopaminergic neurons, rather than promoting general neuronal differentiation. This selectivity is crucial for generating enriched populations of dopaminergic neurons for disease modeling or cell therapy research.

• Neuroprotection and Cell Survival: FGF-20 increases the survival of cultured ventral mesencephalic dopaminergic neurons and protects them against various toxic insults, both in vitro and in vivo. It reduces apoptotic cell death by downregulating markers such as activated caspase-3 and BAX, which is particularly valuable in studies of neurodegeneration and neuroprotection.

• Yield Enhancement in Stem Cell Protocols: When used during the differentiation of hESCs, FGF-20 can increase the yield of tyrosine hydroxylase (TH)-positive dopaminergic neurons up to fivefold, which is significant for generating sufficient cells for downstream applications.

• Disease Modeling and Therapeutic Research: FGF-20’s role in dopaminergic neuron biology makes it a key tool for Parkinson’s disease research, including modeling disease mechanisms, screening neuroprotective compounds, and developing cell replacement therapies.

• Additional Biological Activities: FGF-20 also stimulates proliferation of fibroblasts and is implicated in tissue repair, wound healing, and mucosal restitution, broadening its utility to regenerative medicine and tissue engineering studies.

• Mechanistic Studies: FGF-20 acts via the FGFR-1 receptor, allowing for mechanistic investigations into FGF signaling pathways in neural and non-neural tissues.

In summary, use Recombinant Human FGF-20 in your research if your goals include:

• Generating or protecting dopaminergic neurons from stem cells.
• Investigating neuroprotective mechanisms relevant to Parkinson’s disease.
• Studying FGF signaling in neural development or tissue repair.
• Exploring therapeutic strategies for neurodegeneration or tissue injury.

These properties make FGF-20 a valuable reagent for both basic and translational neuroscience research.

Yes, recombinant human FGF-20 is well-suited for use as a standard for quantification and calibration in ELISA assays. This is actually one of the primary recommended applications for recombinant FGF-20 proteins.

Recommended Formulation and Preparation

When using recombinant FGF-20 as an ELISA standard, you should select the formulation that includes a carrier protein such as bovine serum albumin (BSA). The standard recommendation is to reconstitute the lyophilized protein at 100 μg/mL in sterile phosphate-buffered saline (PBS) containing at least 0.1% human or bovine serum albumin.

Validation and Performance Characteristics

Recombinant FGF-20 standards demonstrate excellent performance in ELISA applications. The calibration curves generated with recombinant FGF-20 standards are typically linear across the detection range, with typical ranges spanning from approximately 15.6 pg/mL to 500 pg/mL or higher, depending on the specific assay kit configuration. The assay sensitivity is generally less than 3.5-3.9 pg/mL, allowing for detection of low FGF-20 concentrations.

Important Consideration

It is critical to note that recombinant FGF-20 proteins formulated specifically as ELISA standards should not be used for bioassay applications, as they are not validated for those purposes. Conversely, if you require the protein for both ELISA standardization and functional bioassays, you should select a carrier-free formulation specifically tested for bioactivity.

The recombinant protein must retain proper tertiary structure for optimal antibody binding and accurate quantification in sandwich ELISA formats. This is why proper reconstitution and storage conditions are essential for maintaining standard integrity throughout your assay work.

Recombinant Human FGF-20 has been validated in published research for several key applications, primarily in functional bioassays, neuroprotection studies, and models of tissue repair and regeneration.

Validated Applications in Published Research:

• Cell Proliferation and Bioactivity Assays:
  Recombinant human FGF-20 is widely used to stimulate proliferation in fibroblast cell lines (e.g., Balb/3T3, NR6R-3T3) as a measure of its biological activity. The ED50 for proliferation in Balb/3T3 cells is typically reported between 10–100 ng/mL.

• Neuroprotection and Dopaminergic Neuron Survival:
  FGF-20 has been shown to enhance the survival of midbrain dopaminergic neurons in vitro and protect against neurotoxic insults (e.g., glutamate, 6-hydroxydopamine) in cell culture and animal models. It has been used in Parkinson’s disease models, where intracerebral infusion of recombinant FGF-20 protects against nigrostriatal tract loss and preserves motor function.

• Blood-Brain Barrier (BBB) Protection:
  In mouse models of traumatic brain injury (TBI), recombinant human FGF-20 has been validated for reducing neurofunctional deficits, brain edema, and neuroinflammation, as well as preserving BBB integrity by upregulating tight and adherens junction proteins.

• Spinal Cord Injury Repair:
  FGF-20 has been used in mouse models to promote spinal cord repair by inhibiting necroptosis, reducing neural deficits, and improving motor function recovery.

• Hair Follicle Biology:
  Recombinant FGF-20 stimulates proliferation and differentiation of hair stromal cells, acting through the Wnt/β-catenin pathway, as shown in dermal papilla cell studies.

• Developmental Biology and Sensory Epithelia:
  FGF-20 is required for sensory epithelial specification in the developing cochlea, validated in mouse models.

• Cancer and Wnt Signaling Studies:
  FGF-20 expression and function have been studied in the context of tumorigenesis, particularly its regulation by β-catenin and its role in oncogenesis via the Wnt pathway.

Summary Table: Key Validated Applications

Additional Notes:

• Most studies use recombinant human FGF-20 in functional assays (e.g., cell proliferation, neuroprotection, tissue repair).
• Applications are typically validated by measuring downstream effects such as cell survival, proliferation, barrier integrity, or functional recovery in animal models.
• The protein is also used to study receptor specificity and signaling pathway activation (e.g., AKT/GSK3β, JNK/NFκB).

If you require protocols or more detailed information on a specific application, please specify the research context.

To reconstitute and prepare Recombinant Human FGF-20 protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA) as a carrier protein. This helps stabilize the protein and prevent adsorption to surfaces.

Step-by-step protocol:

1. Centrifuge the vial briefly (20–30 seconds) before opening to collect all lyophilized powder at the bottom.
2. Add sterile PBS (phosphate-buffered saline) containing at least 0.1% BSA to achieve a final concentration of 100 μg/mL.
   • If your product is carrier-free, you may use sterile PBS alone, but adding BSA is strongly recommended for stability, especially for cell culture applications.
3. Gently mix the solution by pipetting up and down or gentle vortexing until fully dissolved.
4. Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can denature FGF-20.
5. Storage:
   • Store aliquots at −20 °C or below for long-term use.
   • For short-term use (up to 1 week), store at 2–8 °C.
   • Avoid repeated freeze-thaw cycles.

Further dilution:
For cell culture, dilute the stock solution to the desired working concentration (e.g., 10–100 ng/mL) in cell culture medium immediately before use. Always include a carrier protein (such as 0.1% BSA or 10% FBS) in all working solutions to maintain stability.

Additional notes:

• If your protocol or product datasheet specifies a different buffer (e.g., Tris or water), follow those instructions, but PBS with BSA is widely accepted for FGF family proteins.
• For carrier-free preparations, extra care should be taken to minimize protein loss by using low-binding tubes and including a carrier protein in all dilutions.
• Confirm protein solubility visually; if undissolved material remains, gentle mixing or brief sonication may help, but avoid harsh agitation.

Summary Table:

Always consult the specific product datasheet for any unique requirements, as formulations may vary between suppliers.