Recombinant Mouse FGF Acidic

Recombinant Mouse FGF Acidic (FGF1) is widely used in research due to its potent ability to stimulate cell proliferation, promote tissue regeneration, and induce angiogenesis across multiple cell types and tissue origins.

FGF1 is a mitogenic growth factor that acts on cells of mesodermal, neuroectodermal, ectodermal, and endodermal origin, making it valuable for studies involving diverse cell populations. Its key scientific applications include:

• Cell Proliferation and Survival: FGF1 robustly induces DNA synthesis and cell division, supporting expansion of primary cells, stem cells, and cell lines in culture. It also functions as a survival factor by inhibiting p53 activity and pro-apoptotic signaling.
• Angiogenesis: FGF1 is a strong inducer of new blood vessel formation, both in vitro and in vivo, making it essential for vascular biology, wound healing, and cancer research.
• Regeneration and Repair: FGF1 plays a critical role in tissue regeneration, including neural repair, muscle regeneration, and organ development. It has been shown to accelerate nerve regeneration and improve functional recovery after injury in animal models.
• Stem Cell Research: FGF1 supports self-renewal and dedifferentiation, facilitating the maintenance and expansion of pluripotent stem cells and promoting their conversion from differentiated states.
• Developmental Biology: FGF1 is involved in embryogenesis and morphogenesis, regulating processes such as epithelial differentiation and basement membrane metabolism.
• Bioassays and Functional Studies: Recombinant FGF1 is used as a positive control or stimulant in cell-based assays to assess proliferation, migration, and differentiation responses.

Additional technical advantages:

• Species Crossreactivity: Mouse FGF1 shares high sequence identity with human, rat, and other mammalian FGF1, allowing for cross-species experimental designs.
• Defined Activity: Recombinant preparations provide consistent, quantifiable biological activity, essential for reproducible experimental outcomes.

In summary, Recombinant Mouse FGF Acidic is a versatile tool for research in cell biology, regenerative medicine, angiogenesis, and developmental studies, offering robust and reproducible stimulation of cellular processes relevant to tissue growth, repair, and modeling disease mechanisms.

Recombinant Mouse FGF Acidic (FGF1) can be used as a standard for quantification or calibration in ELISA assays, provided it is appropriately validated for your specific assay system.

Key considerations and supporting details:

• Recombinant FGF1 as ELISA Standard: Recombinant Mouse FGF acidic/FGF1 is commonly used as a standard in ELISA assays designed to quantify FGF1 in biological samples. Many commercial ELISA kits for mouse FGF acidic/FGF1 include a recombinant protein standard, and protocols recommend using recombinant FGF1 to generate the standard curve for quantification.

• Validation and Compatibility: It is essential that the recombinant FGF1 you use as a standard is recognized by the antibodies in your ELISA kit. Most well-designed ELISA kits are validated to detect both natural and recombinant forms of FGF1, and their standard curves are generated using recombinant protein. However, you should confirm that your specific ELISA kit documentation states equivalence between natural and recombinant FGF1 detection, as minor differences in post-translational modifications or folding can sometimes affect antibody recognition.

• Carrier Protein Considerations: Recombinant FGF1 is available in carrier-free and carrier-added (e.g., BSA) formulations. Carrier-added forms are generally recommended for use as ELISA standards because they improve protein stability and reduce adsorption to plasticware, leading to more consistent standard curves. If using a carrier-free protein, ensure you add a suitable protein (such as BSA) to your diluent to minimize loss.

• Preparation and Quantification: When preparing your standard curve, reconstitute the recombinant protein according to the manufacturer’s instructions, and use the same diluent as your samples to avoid matrix effects. Prepare serial dilutions covering the expected concentration range in your samples.

• Best Practices:

  • Always run a standard curve in parallel with your samples for each assay plate.
  • Confirm that the standard curve generated with your recombinant FGF1 is linear and covers the dynamic range of your assay.
  • If using a recombinant standard not supplied with your ELISA kit, verify its compatibility by comparing standard curves with those generated using the kit’s original standard, if available.

Limitations:

• If your ELISA kit is designed for a different species or detects only natural FGF1, recombinant mouse FGF1 may not be suitable.
• Some kits or protocols may specify not to use recombinant proteins from other sources due to potential differences in folding or modifications.

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

In summary: You can use recombinant mouse FGF acidic as a standard for ELISA quantification if it is validated for your assay and prepared according to best practices. Always consult your ELISA kit documentation and, if in doubt, perform a parallel comparison with the kit’s supplied standard.

Recombinant Mouse FGF Acidic (FGF1) has been validated for a range of scientific applications in published research, including bioassays, immunohistochemistry, in vivo assays, cell stimulation, and functional studies related to development, regeneration, angiogenesis, and metabolic regulation.

Key validated applications include:

• Bioassays: Used to assess cell proliferation, differentiation, and signaling pathway activation in various cell types, including fibroblasts, endothelial cells, and stem cells.
• Immunohistochemistry: Applied to tissue sections to study FGF1 localization and its role in basement membrane metabolism and epithelial differentiation, particularly in salivary gland and organogenesis studies.
• In Vivo Assays: Employed in mouse models to investigate physiological effects such as skeletal muscle atrophy, diabetes remission, and tissue regeneration.
• Cell Stimulation: Used to stimulate pluripotent stem cells and organoid cultures, supporting studies in kidney organogenesis and retinal differentiation.
• IP (Immunoprecipitation): Utilized to study protein-protein interactions and signaling pathways, especially in tumor vasculature and resistance mechanisms.
• Wound Repair and Tissue Regeneration: Demonstrated efficacy in promoting wound closure, tissue repair, and regeneration in animal models.
• Metabolic Regulation: Central administration of recombinant FGF1 induces sustained diabetes remission in mouse models, highlighting its role in glucose homeostasis and metabolic disease research.
• Angiogenesis: Validated as a potent angiogenic factor, modulating endothelial cell migration and proliferation.
• Developmental Biology: Used to study embryonic development, morphogenesis, and organogenesis, including kidney and salivary gland development.

Sample types validated include:

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

Representative published studies:

• Regulation of salivary gland basement membrane metabolism and epithelial differentiation (Nature Communications, immunohistochemistry).
• Skeletal muscle atrophy and metabolic disease models (International Journal of Molecular Sciences, in vivo assay).
• Stem cell homeostasis and organogenesis (Nature Communications, bioassay; PLoS ONE, stimulation).
• Tumor vasculature and resistance to anti-VEGF therapy (Scientific Reports, IP).
• Retinal and kidney differentiation in stem cell cultures (Stem Cells Translational Medicine, bioassay; PLoS ONE, stimulation).

Summary Table: Validated Applications of Recombinant Mouse FGF Acidic (FGF1)

These applications are supported by multiple peer-reviewed studies and reviews, confirming the broad utility of recombinant mouse FGF acidic in cell biology, developmental biology, regenerative medicine, and disease modeling.

To reconstitute and prepare Recombinant Mouse FGF Acidic (FGF-1) protein for cell culture experiments, follow these best practices based on manufacturer guidelines and scientific recommendations:

Reconstitution

1. Centrifuge the vial: Before opening, briefly centrifuge the lyophilized protein vial in a microcentrifuge for 20–30 seconds to ensure all powder is at the bottom.

2. Reconstitution buffer:

   • Most protocols recommend reconstituting at 100 µg/mL in sterile PBS (phosphate-buffered saline).
   • For improved stability and to prevent adsorption, add 0.1% human or bovine serum albumin (BSA or HSA) to the PBS.
   • Some sources allow reconstitution in sterile distilled water, but PBS with carrier protein is preferred for cell culture applications.

3. Procedure:

   • Add the appropriate volume of sterile PBS (with 0.1% BSA if desired) to achieve a concentration of 100 µg/mL.
   • Gently swirl or invert the vial to dissolve the protein. Avoid vigorous shaking or foaming.
   • Allow the solution to sit at room temperature for 10–15 minutes to ensure complete dissolution.

Preparation for Cell Culture

1. Aliquot and store:

   • Prepare working aliquots to avoid repeated freeze-thaw cycles.
   • Store aliquots at –20°C or –80°C (preferably in a manual defrost freezer).
   • For short-term use (within a few days), aliquots can be kept at 2–8°C.

2. Dilution for cell culture:

   • Dilute the reconstituted stock in cell culture medium or PBS (with or without carrier protein) to the desired working concentration.
   • Typical working concentrations for cell culture range from 1–100 ng/mL, depending on the cell type and experimental design.

3. Addition to cells:

   • Add the diluted FGF-1 directly to the culture medium.
   • Mix gently to ensure even distribution.

Additional Tips

• Avoid repeated freeze-thaw cycles: This can reduce protein activity.
• Use carrier protein: BSA or HSA helps stabilize FGF-1 and prevents loss due to adsorption to plastic surfaces.
• Confirm activity: If possible, run a small test experiment to confirm biological activity before large-scale use.

Example Protocol

• Reconstitute 10 µg lyophilized FGF-1 in 100 µL sterile PBS + 0.1% BSA (final concentration: 100 µg/mL).
• Aliquot 10 µL per tube and store at –20°C.
• For cell culture, dilute 1 µL of stock into 1 mL medium (final concentration: 100 ng/mL).

This approach ensures optimal stability and activity of Recombinant Mouse FGF Acidic for cell culture experiments.