Recombinant Mouse FGF-Acidic

Recombinant mouse FGF-acidic (FGF1) is a valuable tool for research applications due to its potent biological activities and well-characterized functions across multiple cellular and tissue systems.

Key Biological Activities

Recombinant mouse FGF-acidic is a particularly potent inducer of DNA synthesis and possesses strong chemotactic activities. The protein stimulates proliferation across diverse cell types, including all cells of mesodermal origin and many cells of neuroectodermal, ectodermal, and endodermal origin. This broad mitogenic capacity makes it useful for studying cell proliferation mechanisms and for supporting cell culture applications.

Established Research Applications

Tissue Regeneration and Wound Healing

FGF-acidic plays important roles in development, regeneration, and angiogenesis. It has demonstrated efficacy in neural regeneration studies, where application of recombinant FGF-acidic has been shown to facilitate the regeneration of spinal ascending and descending tracts and improve motor behavior recovery following spinal cord injury in animal models. The protein also participates in wound healing applications, with FGF1 showing greater effectiveness than other FGF family members like FGF7 and FGF10.

Cell Culture and Stem Cell Applications

Recombinant mouse FGF-acidic has been used to induce DNA synthesis in cultured cells and supports stem cell maintenance and differentiation protocols. Its non-glycosylated nature and consistent protein composition make it suitable for standardized cell culture experiments.

Signaling Studies

The protein activates well-characterized signaling cascades including phospholipase Cγ (PLCγ), PI3K–AKT, and RAS–MAPK pathways through fibroblast growth factor receptor (FGFR) dimerization and autophosphorylation. This makes it valuable for investigating FGF/FGFR signaling mechanisms in various biological contexts.

Technical Advantages

Recombinant mouse FGF-acidic is typically expressed in bacterial systems as a non-glycosylated protein consisting of 141 amino acid residues with a molecular weight of approximately 15.9–16 kDa. The protein exhibits considerable species cross-reactivity, sharing high amino acid sequence identity with FGF-acidic from other mammalian species, which can inform translational research applications.

Yes, recombinant Mouse FGF-Acidic (FGF1) can generally be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your specific assay system. This is a common practice in ELISA development and quantification of growth factors.

Key considerations and supporting details:

• Assay Compatibility: Most commercial ELISA kits for mouse FGF-acidic/FGF1 are designed to detect both natural and recombinant forms of the protein, and their standard curves are often generated using recombinant FGF1. The assay documentation typically specifies that the kit recognizes both forms and that recombinant protein is suitable for use as a standard.

• Standard Preparation: When preparing a standard curve, it is best practice to use a highly purified recombinant protein, ideally in the same buffer as your samples or as recommended by the kit protocol. If your recombinant FGF1 is carrier-free, ensure it is reconstituted and diluted according to the assay requirements, often with a protein stabilizer such as BSA to prevent adsorption to plasticware.

• Validation: For accurate quantification, confirm that the dose-response curve generated with your recombinant FGF1 is parallel to that of the kit’s standard (if provided), and that the antibodies in the assay bind equivalently to both recombinant and endogenous FGF1. This ensures the assay’s quantification is valid for your samples.

• Source and Purity: The recombinant protein should be of high purity and, if possible, sourced from a reputable supplier or produced under conditions that ensure correct folding and activity. The presence of tags, fusion partners, or contaminants can affect assay performance.

• Carrier Protein: Some recombinant FGF1 preparations contain carrier proteins (e.g., BSA) for stability, which is generally acceptable for ELISA standards unless your assay specifically requires carrier-free protein. If using a carrier-free preparation, add a small amount of BSA or another inert protein to prevent loss due to adsorption.

• Documentation Example: ELISA kit manuals and technical datasheets frequently state: “This assay recognizes natural and recombinant Mouse FGF acidic. No significant cross-reactivity or interference with other proteins was observed”.

Summary Table: Use of Recombinant Mouse FGF-Acidic as ELISA Standard

In summary: Using recombinant Mouse FGF-Acidic as a standard is standard practice for ELISA quantification, provided it is validated for your assay and prepared according to best practices. Always consult your specific ELISA kit documentation for any unique requirements.

Recombinant Mouse FGF-Acidic (FGF1) has been validated in published research for applications including bioassays, immunohistochemistry, in vivo assays, stimulation of cell cultures, and immunoprecipitation.

Key validated applications in peer-reviewed studies:

• Bioassay: Used to assess biological activity, such as stimulation of cell proliferation, differentiation, and survival in various cell types, including mesenchymal stem cells, fibroblasts, and endothelial cells.
• Immunohistochemistry: Applied to tissue sections to study localization and expression patterns of FGF1, particularly in developmental and regenerative contexts (e.g., salivary gland basement membrane metabolism).
• In vivo assay: Employed in live animal models to investigate physiological roles, such as muscle atrophy, stem cell homeostasis, and resistance mechanisms in tumor vasculature.
• Stimulation of cell cultures: Used to induce organogenesis (e.g., kidney organoids from pluripotent stem cells) and differentiation in stem cell protocols.
• Immunoprecipitation (IP): Utilized to study protein-protein interactions and signaling pathways involving FGF1 in cellular extracts.
• Functional assays: Includes assessment of angiogenic activity, wound healing, and DNA synthesis in various cell types.

Additional validated uses:

• ELISA and Western Blot: For detection and quantification of FGF1 in biological samples.
• Cell migration and proliferation assays: To study FGF1’s role as a mitogen and angiogenic factor.

Representative published studies:

• Regulation of mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis (Nature Communications, 2023).
• Role in salivary gland morphogenesis and basement membrane metabolism (Nature Communications, J. Biol. Chem.).
• Investigation of skeletal muscle atrophy mechanisms in mice (Int. J. Mol. Sci.).
• Kidney organogenesis from pluripotent stem cells (PLoS ONE).
• Tumor vasculature resistance to anti-VEGF therapy (Sci Rep).

Summary of sample types:

• Whole cells
• Whole tissue
• Cell culture supernatants
• In vivo animal models

Scientific context:
FGF1 is widely used in research on development, regeneration, angiogenesis, stem cell biology, and disease models due to its potent mitogenic and survival-promoting properties. Its applications span both basic mechanistic studies and translational research, including tissue engineering and regenerative medicine.

If you require protocol details or specific experimental setups for any of these applications, please specify the intended use.

To reconstitute and prepare Recombinant Mouse FGF-Acidic (FGF1) protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile phosphate-buffered saline (PBS). For enhanced stability, especially during storage or repeated freeze-thaw cycles, include 0.1% bovine or human serum albumin (BSA/HSA) as a carrier protein.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Add sterile PBS (pH 7.4) to achieve a final concentration of 100 μg/mL. If recommended by your supplier or for long-term storage, use PBS containing 0.1% BSA/HSA.
• Gently pipette the solution down the sides of the vial to dissolve the protein. Do not vortex, as vigorous mixing can denature the protein.
• Allow the protein to fully dissolve at room temperature for several minutes.
• Once dissolved, aliquot the solution to avoid repeated freeze-thaw cycles.
• Store aliquots at -20°C to -70°C for up to 3 months, or at 2–8°C for up to 1 month under sterile conditions.
• For cell culture, further dilute the stock solution in your culture medium immediately before use. If your application is sensitive to carrier proteins, ensure your final working solution is compatible with your assay.

Additional notes:

• If your protocol or supplier specifies, you may use sterile distilled water for initial reconstitution, but PBS is generally preferred for physiological compatibility.
• Always consult the product’s Certificate of Analysis or datasheet for specific instructions, as formulations may vary.
• Avoid repeated freeze-thaw cycles by preparing single-use aliquots.
• For bioactivity, ensure the final working concentration is optimized for your cell type and experimental design.

Summary Table:

This protocol ensures optimal solubility, stability, and bioactivity of recombinant mouse FGF-acidic protein for cell culture applications.