Recombinant Mouse FGF-8c

Recombinant Mouse FGF-8c is used in research applications to study and manipulate developmental processes, particularly those involving epithelial-mesenchymal interactions, cell proliferation, differentiation, and tissue patterning during embryogenesis. It is especially valuable for dissecting the specific signaling pathways mediated by FGF-8c and its receptor specificity.

Key reasons to use Recombinant Mouse FGF-8c in research:

• Isoform-Specific Receptor Activation: FGF-8c selectively activates the 'c' splice form of FGFR3 and FGFR4, which are predominantly expressed in mesenchymal tissues during development. This allows researchers to study isoform- and receptor-specific signaling events, which is critical for understanding tissue patterning and organogenesis.

• Developmental Biology Applications: FGF-8c is highly expressed in regions of active morphogenesis, such as the developing limb and craniofacial structures, and plays a central role in processes like gastrulation, somitogenesis, and morphogenesis. Using recombinant FGF-8c enables controlled studies of these developmental events in vitro and in vivo.

• Epithelial-Mesenchymal Signaling: FGF-8c, produced by epithelial cells, provides mitogenic signals to underlying mesenchyme, making it essential for modeling epithelial-mesenchymal interactions in organoid cultures, tissue engineering, and regenerative medicine.

• High Purity and Consistency: Recombinant proteins offer high bioactivity, batch-to-batch consistency, and purity, which are crucial for reproducible experimental outcomes, especially in organoid and stem cell research.

• Disease Modeling and Cancer Research: Overexpression of FGF-8 isoforms, including FGF-8c, has been linked to increased tumor growth and angiogenesis, making it a useful tool for cancer biology studies and for investigating mechanisms of tumor progression and vascularization.

• Functional Studies: Recombinant FGF-8c can be used to stimulate cell proliferation, differentiation, and migration in various cell types, facilitating studies on tissue repair, regeneration, and developmental defects.

In summary, Recombinant Mouse FGF-8c is a powerful tool for developmental biology, disease modeling, and tissue engineering due to its defined receptor specificity, role in key morphogenetic processes, and the experimental advantages of recombinant protein technology.

Recombinant Mouse FGF-8c can be used as a standard for quantification or calibration in ELISA assays, provided the ELISA is validated to recognize recombinant FGF-8c and the standard curve is prepared appropriately.

Key considerations and supporting details:

• ELISA Kit Specificity: Some ELISA kits are designed to detect only the native (endogenous) form of FGF-8 and may not recognize recombinant proteins due to differences in folding, post-translational modifications, or epitopes. Always check the kit documentation to confirm whether it detects recombinant FGF-8c.

• Parallelism and Validation: For quantitative ELISA, the standard (recombinant FGF-8c) must produce a dose-response curve that is parallel to that of the endogenous protein in your sample matrix. This ensures that the assay quantifies both forms equivalently. If the curves are not parallel, quantification may be inaccurate.

• Standard Preparation: Recombinant proteins are commonly used as standards in ELISA assays, especially when purified native protein is unavailable. The standard should be reconstituted and diluted according to the ELISA kit instructions to generate a standard curve covering the expected concentration range.

• Carrier Proteins: Recombinant proteins may be supplied with or without carrier proteins (e.g., BSA). For ELISA standards, carrier-free preparations are often preferred to avoid interference, unless the kit specifically recommends otherwise.

• Documentation Examples: Some ELISA kits for other FGF family members (e.g., FGF-21, FGF-basic/FGF2) explicitly state that recombinant proteins are suitable as standards and that the assay quantifies both recombinant and natural forms accurately. For FGF-8, kit protocols typically instruct users to prepare standards from recombinant protein.

• Best Practice: Always validate the use of recombinant FGF-8c as a standard in your specific ELISA system by:

  • Running a standard curve with the recombinant protein.
  • Testing for parallelism with endogenous FGF-8 in your sample matrix.
  • Confirming that the assay’s antibodies recognize the recombinant form.

If your ELISA kit documentation or technical support confirms compatibility with recombinant FGF-8c, and you validate parallelism, you can confidently use it as a standard for quantification or calibration in your assays.

Recombinant Mouse FGF-8c has been validated primarily for use in bioassays in published research, particularly in studies involving cell signaling, neurogenesis, embryonic development, and cell differentiation in mouse models. Most published applications focus on its role as a signaling molecule in cell-based and tissue culture systems.

Key validated applications include:

• Bioassays: FGF-8c is widely used to stimulate or modulate cellular responses in vitro, such as proliferation, differentiation, and survival of various cell types, including neural progenitors, embryonic stem cells, and other primary or immortalized cell lines.
• Neurogenesis and Neural Differentiation: Studies have used recombinant FGF-8c to investigate its effects on neural stem/progenitor cell fate, regional patterning of the developing brain, and modulation of gene expression relevant to neurodevelopment.
• Embryonic Development: FGF-8c has been applied in assays modeling embryonic tissue patterning, limb development, and organogenesis, reflecting its physiological roles in vivo.
• Cell Proliferation and Migration: It is used to assess its impact on cell proliferation and migration, especially in developmental and regenerative contexts.
• Disease Models: Some studies have used FGF-8c in mouse models to explore its involvement in disease processes, such as neurodegeneration or tumorigenesis, by modulating relevant signaling pathways.

Experimental formats where FGF-8c has been validated:

• Whole cell cultures (primary cells, stem cells, immortalized lines).
• 3D organoid systems for modeling brain development and regionalization.
• In vivo mouse models for developmental biology and disease research.

Assay types:

• Cell proliferation assays (e.g., NR6R-3T3 cell proliferation).
• Gene expression analysis (qRT-PCR, single-cell RNA-seq).
• Immunostaining for neural and developmental markers.
• Functional assays for differentiation and tissue patterning.

Summary Table: Validated Applications of Recombinant Mouse FGF-8c

No published research was found validating FGF-8c for immunoassays (e.g., ELISA), flow cytometry, or western blot as a standard or detection reagent; its use is focused on functional and mechanistic cell-based assays.

If you require details on a specific application or protocol, please specify the context (e.g., neural differentiation, organoid culture, in vivo developmental studies).

To reconstitute and prepare Recombinant Mouse FGF-8c protein for cell culture experiments, follow these steps:

• Centrifuge the vial briefly (20–30 seconds) before opening to ensure all lyophilized protein is at the bottom.
• Reconstitute the protein in sterile distilled water or an aqueous buffer containing 0.1% BSA (bovine serum albumin) to a final concentration of 0.1–1.0 mg/mL. A commonly used concentration is 0.1 mg/mL.
• Gently pipette to dissolve the protein, washing down the sides of the vial to maximize recovery.
• Allow several minutes for complete dissolution.

Best practices for cell culture use:

• After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can denature the protein.
• For short-term storage (up to 1 month), keep aliquots at 2–8 °C under sterile conditions.
• For long-term storage (up to 3 months), store at –20 °C to –70 °C.
• If using a carrier protein (such as BSA), ensure it is present at 0.1% to stabilize the protein and prevent adsorption to tube walls.
• Before use in cell culture, further dilute the reconstituted stock in your cell culture medium as required for your assay.

Summary protocol:

1. Briefly centrifuge the vial.
2. Add sterile distilled water or buffer with 0.1% BSA to achieve 0.1–1.0 mg/mL.
3. Gently mix and allow to dissolve completely.
4. Aliquot and store at recommended temperatures.
5. Avoid repeated freeze-thaw cycles.

Note: Always consult the specific product datasheet or certificate of analysis for any manufacturer-specific instructions, as minor formulation differences may exist.