Recombinant Human FGF-Acidic

Recombinant Human FGF-Acidic (FGF-1) is widely used in research due to its potent ability to stimulate cell proliferation, promote tissue regeneration, and drive angiogenesis, making it valuable for studies in development, wound healing, and disease modeling.

Key scientific reasons to use recombinant human FGF-acidic in research applications:

• Potent Mitogen: FGF-1 is a strong activator of DNA synthesis and cell proliferation across a broad range of cell types, including fibroblasts, endothelial cells, and cells of mesodermal and neuroectodermal origin.
• Angiogenesis and Tissue Repair: It plays a central role in angiogenesis (formation of new blood vessels), tissue regeneration, and wound healing, making it essential for studies in regenerative medicine and tissue engineering.
• Developmental Biology: FGF-1 is involved in organ development (e.g., skin, brain, muscle, bone, heart) and is critical for studying developmental processes and morphogenesis.
• Cell Survival and Differentiation: It supports cell survival, differentiation, and migration, which are important for stem cell research, neuronal studies, and modeling of various physiological and pathological processes.
• Disease Modeling: FGF-1 is implicated in cancer biology (promoting tumor cell proliferation and survival), neuroprotection, and metabolic regulation, making it useful for disease modeling and therapeutic research.
• Cross-Species Reactivity: Recombinant human FGF-1 is highly conserved and active across multiple species, facilitating translational studies and comparative biology.
• Defined, Reproducible Reagent: Using recombinant protein ensures batch-to-batch consistency, purity, and the absence of animal-derived contaminants, which is critical for reproducibility and regulatory compliance in research and preclinical studies.

Common research applications include:

• Enhancing proliferation and maintenance of stem cells in culture.
• Promoting wound healing and tissue regeneration in in vitro and in vivo models.
• Studying angiogenesis and vascular biology.
• Investigating mechanisms of cell signaling, differentiation, and survival.
• Modeling cancer progression and testing anti-angiogenic or anti-proliferative therapies.
• Exploring neuroprotection and neuronal differentiation.

In summary, recombinant human FGF-acidic is a versatile and well-characterized growth factor that enables precise control of cellular processes in a wide range of experimental systems, supporting both basic and translational research.

Yes, recombinant human FGF-acidic (FGF1) can be used as a standard for quantification or calibration in ELISA assays, provided it is formulated and validated for this purpose. Recombinant FGF1 is commonly used as a standard in commercial ELISA kits designed to measure human FGF-acidic in biological samples.

Key considerations for use as an ELISA standard:

• Formulation: Recombinant FGF1 intended for use as an ELISA standard is often supplied with a carrier protein such as BSA to enhance stability and prevent adsorption to plasticware. Carrier-free formulations are available but may require additional precautions to maintain protein stability and prevent loss due to adsorption.

• Validation: The recombinant protein should be validated for use as a standard in ELISA. Commercial ELISA kits typically use E. coli-expressed recombinant human FGF-acidic as the calibration standard and have demonstrated accurate quantitation of the recombinant factor. It is important to ensure that the recombinant standard matches the analyte detected by the antibodies in your assay (e.g., same isoform, sequence, and post-translational modifications if relevant).

• Reconstitution and Handling: Follow the manufacturer’s instructions for reconstitution, dilution, and storage. For example, reconstituting at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin is recommended to maintain stability.

• Assay Matrix: When preparing standard curves, dilute the recombinant standard in the same buffer or matrix as your samples to minimize matrix effects and ensure accurate quantification.

• Precision and Range: Commercial ELISA kits using recombinant FGF-acidic as a standard report high intra- and inter-assay precision (CV <10%) and a typical detection range from ~10–1000 pg/mL, depending on the kit.

Limitations and Best Practices:

• Bioactivity vs. Immunoreactivity: While recombinant FGF1 may be biologically active, for ELISA calibration only immunoreactivity (antibody recognition) is required. Ensure the recombinant standard is recognized by the antibodies used in your assay.

• Carrier Protein Interference: If your assay is sensitive to carrier proteins (e.g., BSA), use carrier-free recombinant FGF1 and take precautions to prevent protein loss due to adsorption.

• Source and Sequence: Confirm that the recombinant FGF1 standard is full-length and matches the sequence detected by your ELISA antibodies (e.g., aa 16–155 for human FGF-acidic).

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

In conclusion, recombinant human FGF-acidic is suitable as a standard for ELISA quantification if properly formulated, validated, and handled according to best practices.

Recombinant Human FGF-Acidic (FGF1) has been validated in published research for a range of applications, primarily as a bioactive growth factor in cell-based assays, cell culture, and functional studies.

Key validated applications include:

• Bioassays: FGF1 is widely used to assess its mitogenic (cell proliferation-inducing) activity in various cell types, including fibroblasts, endothelial cells, and stem cells. These assays often measure DNA synthesis, cell proliferation, or survival in response to FGF1 stimulation.

• Cell Culture Supplementation: FGF1 is used to support the growth and maintenance of stem cells, including human embryonic stem cells, induced pluripotent stem cells (iPSCs), and differentiation protocols toward specific lineages such as hepatocytes and photoreceptors. It is also used in xeno-free culture systems for retinal differentiation and other regenerative medicine applications.

• Functional Studies: Published research has validated FGF1 for:

  • Neuroprotection and neuronal differentiation: FGF1 promotes neuronal survival and differentiation, and is involved in studies of neurodegeneration and brain injury.
  • Wound healing and tissue regeneration: FGF1 is used in models of tissue repair, including skin and liver regeneration, due to its role in angiogenesis and cell migration.
  • Angiogenesis assays: FGF1 is a potent inducer of angiogenesis, validated in both in vitro and in vivo models.
  • Oxidative stress and inflammation: Studies have used FGF1 to investigate its protective effects against oxidative and endoplasmic reticulum stress, particularly in retinal pigment epithelial cells under diabetic conditions.
  • Cancer research: FGF1 is used to study tumor cell proliferation, tumor microenvironment interactions, and as a target for cancer immunotherapy.

• ELISA, Western Blot, and Immunohistochemistry: While less common than functional assays, FGF1 has been validated as a standard or control in ELISA, Western blot, and immunohistochemistry protocols.

• Enzymatic Activity Assays: FGF1 has been used in vitro to assess its enzymatic or signaling activity, such as stimulation of thymidine uptake or activation of downstream pathways.

Summary Table of Validated Applications

Species and Sample Types: FGF1 has been validated in human, mouse, rat, and canine cells, and is reactive with a broad range of species.

References to Published Research: Numerous peer-reviewed studies have used recombinant human FGF1 for these applications, including investigations into retinal protection, stem cell differentiation, tumor biology, and tissue engineering.

Note: FGF1 is distinct from FGF2 (basic FGF), though both share overlapping applications. Always confirm the specific isoform and application in your experimental context.

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

Reconstitution

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

2. Choose the reconstitution buffer:

   • Most protocols recommend sterile PBS (pH 7.4) or sterile water.
   • Some suppliers suggest 5 mM sodium phosphate or 5–10 mM Tris (pH 7.6).
   • For maximum stability, use a buffer containing at least 0.1% BSA (bovine serum albumin) or HSA (human serum albumin) to minimize protein loss due to adsorption.

3. Reconstitution concentration:

   • Typical reconstitution concentrations are 100–200 µg/mL (0.1–0.2 mg/mL).
   • For example, dissolve 50 µg of lyophilized protein in 250–500 µL of buffer.

4. Gentle mixing:

   • Gently swirl or pipette up and down to dissolve the protein. Avoid vigorous shaking to prevent denaturation.

Preparation for Cell Culture

1. Aliquot and store:

   • Aliquot the reconstituted protein into small volumes and store at –20°C (or –80°C for longer storage).
   • Avoid repeated freeze-thaw cycles to maintain activity.

2. Prepare working dilutions:

   • For cell culture, dilute the stock solution in appropriate cell culture medium or buffer containing 0.1% BSA or serum to minimize protein loss.
   • Prepare dilutions immediately before use.

3. Storage of working solutions:

   • Use diluted solutions within a few hours or store at 4°C for up to 1 month if necessary.
   • For longer storage, aliquot and freeze at –20°C or –80°C.

Additional Tips

• Avoid frost-free freezers for storage, as temperature fluctuations can degrade the protein.
• If a precipitate forms, microcentrifuge briefly before use.
• For sensitive assays, confirm protein concentration and activity using a suitable assay (e.g., ELISA or cell proliferation assay).

Example Protocol

1. Centrifuge vial (20–30 sec).
2. Reconstitute 50 µg protein in 250 µL sterile PBS + 0.1% BSA (final concentration: 200 µg/mL).
3. Aliquot into small tubes and freeze at –20°C.
4. For cell culture, dilute to desired concentration (e.g., 10–100 ng/mL) in medium containing 0.1% BSA or serum.
5. Use immediately or store diluted solution at 4°C for up to 1 month.

Following these steps will help ensure optimal activity and stability of Recombinant Human FGF-Acidic for your cell culture experiments.