Recombinant Human FGF-Basic

Recombinant Human FGF-Basic (FGF-2/bFGF) is a versatile growth factor that offers significant advantages across multiple research and therapeutic applications due to its broad biological activity and well-characterized mechanisms of action.

Cellular Proliferation and Differentiation

FGF-Basic possesses broad mitogenic activity and stimulates proliferation across diverse cell types, including fibroblasts, myoblasts, osteoblasts, neuronal cells, endothelial cells, keratinocytes, chondrocytes, astrocytes, oligodendrocytes, and smooth muscle cells. Beyond proliferation, it induces neuronal differentiation, survival, and regeneration, making it particularly valuable for studies involving neural development and function. The protein also significantly promotes the proliferation of adipose-derived mesenchymal cells and enhances chondrogenesis in three-dimensional micromass culture.

Tissue Regeneration and Wound Healing

FGF-Basic demonstrates remarkable effectiveness in promoting tissue regeneration through multiple complementary mechanisms. It maintains osteoblast precursors in a proliferative state while simultaneously promoting angiogenesis, which is crucial for nutrient and oxygen supply in newly formed tissue. The protein stimulates secretion of other growth factors, such as vascular endothelial growth factor and hepatocyte growth factor, further enhancing vascular development and creating a favorable microenvironment for regeneration. In bone regeneration applications specifically, optimal efficacy has been demonstrated at a 0.3% concentration, which has received formal regulatory approval for periodontal regenerative medicine. Chronic wound research represents one of the largest areas of FGF-2 investigation, as these wounds often have reduced endogenous FGF-2 concentrations.

Angiogenic Activity

FGF-Basic exhibits potent angiogenic activity and plays a key role in both physiological and pathological conditions. This property makes it essential for studying vascular development, wound repair, inflammation, and tumor biology. The protein binds to a family of four distinct, high-affinity tyrosine kinase receptors (FGFR-1 to FGFR-4) and also interacts with αvβ3 integrin, enabling multiple signaling pathways.

Diverse Research Applications

The protein's versatility extends to numerous research contexts, including embryonic development, stem cell maintenance, organoid culture, and disease modeling. It has proven valuable in bioassay applications across whole cell systems and organoid models, supporting investigations ranging from basic cell biology to translational research.

Yes, recombinant human FGF-basic (FGF2) can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system. Multiple sources confirm that recombinant FGF-basic is routinely used as a standard in commercial ELISA kits and for assay calibration.

Key considerations and supporting details:

• Assay Compatibility:
  Recombinant human FGF-basic is commonly used as a standard in ELISA kits designed to quantify FGF2 in biological samples. These kits are validated to detect both natural and recombinant forms, and standard curves generated with recombinant FGF-basic are used for quantification.

• Protein Form and Source:
  The recombinant protein is typically expressed in E. coli and is available in carrier-free or BSA-containing formulations. Carrier-free forms are preferred if BSA or other additives could interfere with your assay.

• Standard Curve Validity:
  Commercial ELISA kits demonstrate that standard curves generated with recombinant FGF-basic are linear and parallel to those generated with natural FGF-basic, indicating equivalence for quantification purposes. This ensures accurate calibration and quantification of FGF2 in your samples.

• Reconstitution and Handling:
  Follow the manufacturer’s instructions for reconstitution (e.g., using Tris buffer, gentle mixing) and prepare further dilutions in a buffer containing a carrier protein (such as 0.1% BSA) to minimize protein loss.

• Stability:
  Recombinant FGF-basic can be unstable in aqueous solution, so aliquot and store at recommended temperatures, and avoid repeated freeze-thaw cycles to maintain protein integrity and assay accuracy.

• Concentration Range:
  ELISA kits using recombinant FGF-basic as a standard typically cover a wide dynamic range (from low pg/mL to ng/mL), suitable for most biological sample quantification needs.

Best Practices:

• Confirm that your ELISA antibodies recognize the recombinant form you are using (e.g., full-length, specific isoform, or tag-free).
• Validate the standard curve in your specific assay matrix (serum, plasma, cell culture supernatant) for optimal accuracy.
• Use freshly prepared or properly stored aliquots of the recombinant standard to ensure consistent results.

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

In conclusion:
Recombinant human FGF-basic is suitable and widely accepted as a standard for quantification and calibration in ELISA assays, provided it matches the requirements of your specific assay system and is handled according to best practices.

Recombinant Human FGF-Basic (FGF2/bFGF) has been validated in published research for a wide range of applications, primarily in cell culture, bioassays, tissue engineering, regenerative medicine, and therapeutic studies.

Key validated applications include:

• Stem Cell Culture and Maintenance:
  FGF-basic is widely used to maintain the proliferation and pluripotency of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs), supporting their undifferentiated state and self-renewal. It upregulates pluripotency markers such as Oct4, Sox2, and Nanog.

• Cell Differentiation and Developmental Biology:
  It is used to induce differentiation of various cell types, including neuronal cells, Schwann cells, chondrocytes, and fibroblasts, and to study embryonic development and neurogenesis. FGF-basic has been applied in protocols for deriving neural crest cells and Schwann cells from hPSCs for disease modeling.

• Angiogenesis and Wound Healing:
  FGF-basic promotes angiogenesis, tissue repair, and regeneration, and is validated for use in wound healing models, including clinical studies for burn treatment and diabetic wound healing. It accelerates skin cell regeneration and improves microcirculation.

• Bioassays and Functional Studies:
  Used in bioassays to assess cell proliferation, survival, and differentiation in various cell types, including endothelial cells, neuronal cells, and mesenchymal stem cells. It is also applied in functional assays such as ELISA standards.

• Cancer Research:
  FGF-basic is implicated in tumor angiogenesis and cell proliferation, and is used in studies investigating its role in cancer development and tumor vascularization.

• Tissue Engineering and Regenerative Medicine:
  Supplementation of culture media with FGF-basic supports the growth and expansion of chondrocyte-like cells and other progenitor cells for tissue engineering applications.

• Genome Editing and Organoid Models:
  FGF-basic is used in protocols for high-precision genome editing (HDR) and in the culture of organoids for disease modeling and drug discovery.

• Therapeutic Applications:
  Clinical studies have validated recombinant human FGF-basic for therapeutic use in wound healing, particularly in burn patients and diabetic ulcers, demonstrating improved healing rates and safety.

Summary Table of Validated Applications

Additional Notes:

• FGF-basic is validated for use with a variety of cell types, including fibroblasts, endothelial cells, neuronal cells, chondrocytes, and stem cells.
• It is commonly used in both in vitro (cell culture, organoid) and in vivo (animal models, clinical studies) settings.
• Protocols often specify carrier-free or animal-free formulations for clinical and stem cell applications.

If you require protocol details or specific assay conditions for any application, please specify the intended use.

To reconstitute and prepare Recombinant Human FGF-Basic (FGF-2/bFGF) protein for cell culture experiments, follow these best-practice steps to ensure protein stability and bioactivity:

1. Preparation Before Reconstitution

• Briefly centrifuge the vial (10,000–14,000 × g, 20–30 seconds) to collect all lyophilized protein at the bottom and cap of the vial.

2. Reconstitution

• Use a sterile, low-protein-binding pipette tip to add the appropriate volume of buffer.
• Recommended buffers:
  • Sterile 1× PBS (pH 7.2–7.4)
  • Tris buffer (5–10 mM, pH 7.5–7.6)
• Protein concentration: Reconstitute to a final concentration of 0.1–0.3 mg/mL (100–300 μg/mL) for stock solutions.
• Carrier protein: Add 0.1% BSA or recombinant human serum albumin (HSA) to the buffer to stabilize the protein and prevent adsorption to surfaces.
• Gently swirl or tap the vial to dissolve the protein. Avoid vigorous vortexing to prevent denaturation.

3. Aliquoting and Storage

• After complete dissolution, aliquot the stock solution into single-use volumes to avoid repeated freeze-thaw cycles.
• Store aliquots at –20 °C or –80 °C in a manual defrost freezer. Avoid frost-free freezers and repeated freeze-thaw cycles, as these can denature the protein.
• For short-term use (up to 1 week), aliquots can be stored at 4 °C.

4. Working Solution Preparation

• Dilute the stock solution to the desired working concentration (typically 10–100 ng/mL for most cell culture applications) in cell culture medium immediately before use.
• Include 0.1% BSA or HSA in the working solution if possible to maintain stability.

5. Additional Notes

• If a precipitate forms after reconstitution, clarify by microcentrifugation before use.
• Avoid prolonged storage of reconstituted protein at 4 °C or room temperature, as FGF-Basic is unstable in aqueous solution without stabilizers.
• Confirm protein integrity by SDS-PAGE if needed.

Summary Table: Key Parameters for FGF-Basic Reconstitution

These guidelines will help ensure maximum stability and biological activity of recombinant human FGF-Basic for cell culture experiments.