Recombinant Human FGF-17

Recombinant Human FGF-17 is used in research applications to study and manipulate processes such as embryonic development, neurogenesis, angiogenesis, cell growth, and tissue repair, due to its role as a member of the fibroblast growth factor (FGF) family.

FGF-17 is particularly important for:

• Central nervous system development: It is prominently expressed in the cerebellum and cortex, and is involved in patterning at the midbrain/hindbrain junction during embryogenesis.
• Cell proliferation and survival: Like other FGFs, FGF-17 exhibits broad mitogenic (cell division-promoting) and cell survival activities, making it valuable for studies on cell growth, morphogenesis, and tissue regeneration.
• Angiogenesis and tissue repair: FGFs regulate blood vessel formation and tissue repair, which is relevant for wound healing and regenerative medicine applications.
• Disease modeling: Altered FGF-17 expression is associated with developmental disorders such as Dandy-Walker cerebellar malformation, and knockout models show neurological and behavioral abnormalities, making it useful for disease mechanism studies.

Additional advantages of using recombinant human FGF-17 include:

• Defined activity and purity: Recombinant production ensures consistent biological activity and purity, which is critical for reproducible experimental results.
• Human cell-expressed proteins: If produced in human cells, recombinant FGF-17 offers native folding, glycosylation, and higher stability and activity compared to proteins from non-human systems.

Typical research applications include:

• Neural development and differentiation assays
• Stem cell culture and expansion
• Angiogenesis and vascular biology studies
• Tissue engineering and regenerative medicine
• Disease modeling for neurodevelopmental and vascular disorders

In summary, recombinant human FGF-17 is a versatile tool for investigating developmental biology, cell signaling, and tissue regeneration, with particular relevance to the nervous system and vascular development.

Yes, recombinant human FGF-17 can generally be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity, properly quantified, and compatible with your assay system. Many commercial ELISA kits for FGF-17 use recombinant human FGF-17 as the standard, and the assay antibodies are typically validated to recognize both natural and recombinant forms.

Key considerations:

• Antibody Specificity: Most FGF-17 ELISA kits are designed to detect both natural and recombinant human FGF-17, and their standard curves are often generated using recombinant protein. This ensures that the recombinant standard is appropriate for quantifying FGF-17 in biological samples.

• Protein Formulation: Recombinant FGF-17 is available in different formulations (with or without carrier proteins such as BSA). For ELISA calibration, both carrier-free and BSA-containing forms can be used, but carrier-free is preferred if BSA may interfere with your assay.

• Standard Preparation: Prepare the standard curve by serially diluting the recombinant FGF-17 in the same buffer or matrix as your samples to minimize matrix effects. Always generate a fresh standard curve for each assay run.

• Validation: Ensure that the recombinant FGF-17 you use is of high purity and accurately quantified. If using a recombinant standard not provided with your ELISA kit, it is advisable to validate its performance by comparing its standard curve to that of the kit-provided standard, if available.

• Documentation: Check your ELISA kit manual for any specific recommendations or restrictions regarding the use of external standards. Some kits may require the use of their supplied standards for optimal accuracy.

Summary Table: Use of Recombinant FGF-17 as ELISA Standard

In summary: Using recombinant human FGF-17 as a standard is standard practice for ELISA quantification, provided the protein is compatible with your assay and properly validated.

Recombinant Human FGF-17 has been validated for several key applications in published research, including ELISA, functional (bioactivity) assays, Western blot, blocking assays, immunohistochemistry, and SDS-PAGE.

FGF-17 is a member of the fibroblast growth factor family and is widely used to study its biological roles in angiogenesis, mitogenesis, cell differentiation, tissue repair, and tumor biology. Specific validated applications include:

• ELISA: Used as a standard or analyte to quantify FGF-17 levels in biological samples.
• Functional/Bioactivity Assays: Assessed for its ability to stimulate cell proliferation, survival, and differentiation, often in the presence of heparin. For example, FGF-17 has been shown to induce mitogenic responses in cell lines and primary cells.
• Western Blot: Used as a positive control or to validate antibody specificity for FGF-17 detection.
• Blocking Assays: Employed to study receptor-ligand interactions and downstream signaling by inhibiting FGF-17 activity.
• Immunohistochemistry: Used to localize FGF-17 expression in tissue sections.
• SDS-PAGE: Validated for purity assessment and molecular weight determination.

Research Applications:

• Developmental Biology: FGF-17 is critical for patterning at the midbrain/hindbrain junction and is expressed in developing neural, skeletal, and vascular tissues.
• Cancer Research: Altered FGF-17 expression is associated with prostate cancer, leukemic cell lines, and other malignancies. It is used to study tumor growth, invasion, and FGFR signaling.
• Neuroscience: FGF-17 is prominently expressed in the cerebellum and cortex, with roles in neural development and disease models.
• Tissue Repair and Regeneration: FGF-17 and related FGFs are used to promote cell migration, proliferation, and wound healing in vitro and in vivo models.

Summary Table: Validated Applications for Recombinant Human FGF-17

FGF-17 is also used as a protein standard, immunogen, and in cell culture for various cell biology applications. Its broad validation supports its use in both basic and translational research focused on growth factor signaling, development, and disease.

To reconstitute and prepare Recombinant Human FGF-17 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer—typically sterile PBS or 5 mM Tris, pH 8.0—to a concentration between 0.1–1.0 mg/mL. For enhanced stability and to prevent adsorption, include 0.1% BSA (bovine serum albumin) if the protein is not already formulated with a carrier.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Add sterile buffer (PBS or 5 mM Tris, pH 8.0) to achieve the desired concentration (e.g., for 100 µg protein, add 100 µL for 1 mg/mL or 1 mL for 0.1 mg/mL).
• If the protein is carrier-free, add at least 0.1% BSA to the buffer to stabilize the protein and prevent loss due to adsorption.
• Gently mix the solution to fully dissolve the protein; avoid vigorous vortexing to prevent denaturation.
• If any precipitate remains, microcentrifuge briefly and use the supernatant.
• Aliquot the solution to avoid repeated freeze-thaw cycles, which can denature FGF-17.
• Store aliquots at −20°C to −70°C in a manual defrost freezer for long-term storage, or at 2–8°C for short-term use (up to one month).

Additional notes:

• For cell culture, always use sterile technique and sterile buffers.
• The optimal working concentration for cell culture assays should be determined empirically, but initial reconstitution at 0.1–1.0 mg/mL allows for flexible dilution.
• If the product datasheet or Certificate of Analysis (COA) provides specific instructions, follow those recommendations for buffer composition and concentration.

Summary Table: Reconstitution Options

Careful reconstitution and handling are essential for maintaining the biological activity of FGF-17 in cell culture applications.