Recombinant Human HGF (NS0 Cell Expressed) is widely used in research applications due to its high bioactivity, batch-to-batch consistency, and suitability for cell-based assays, especially those investigating cell proliferation, migration, morphogenesis, and tissue regeneration.

Key scientific advantages and applications include:

• High Bioactivity and Consistency: Recombinant HGF produced in NS0 cells is validated for inducing IL-11 secretion in Saos-2 human osteosarcoma cells, with an ED₅₀ ≤ 4.00 ng/mL, demonstrating potent biological activity and reliable performance across batches.
• Defined Structure and Purity: NS0-expressed HGF is typically supplied as a cleaved, bioactive disulfide-linked heterodimer (α and β chains), closely mimicking the native human protein and minimizing contaminants or unwanted isoforms.
• Relevance to Human Biology: HGF is a key regulator of epithelial morphogenesis, cell scattering, branching tubulogenesis, and upregulation of integrin α2β1, making it essential for studies on organ development, wound healing, cancer biology, and stem cell migration.
• Validated for Multiple Applications: NS0-expressed HGF is suitable for bioassays, cell culture, binding studies, and organoid research, supporting experiments in tissue engineering, regenerative medicine, and oncology.
• Reduced Animal Component Contamination: Recombinant production in NS0 (murine myeloma) cells avoids animal-derived contaminants common in native preparations, supporting reproducibility and regulatory compliance for translational research.

Typical research uses include:

• Stimulating cell proliferation, motility, and differentiation in epithelial, endothelial, and stem cells.
• Modeling tissue regeneration and wound healing processes.
• Investigating MET receptor signaling in cancer and developmental biology.
• Supporting organoid and 3D culture systems with defined growth factor supplementation.

In summary, Recombinant Human HGF (NS0 Cell Expressed) is preferred for its robust activity, purity, and reliability in diverse cell-based and molecular assays, making it a critical reagent for studies of cell signaling, tissue morphogenesis, and regenerative processes.

Yes, you can use recombinant Human HGF (NS0 cell expressed) as a standard for quantification or calibration in ELISA assays, provided it is formulated appropriately for this purpose. Recombinant HGF expressed in NS0 cells is widely used as a calibrator in commercial ELISA kits and is recognized for its parallelism with natural human HGF in assay standard curves.

Key considerations and best practices:

• Formulation: For use as an ELISA standard, it is generally recommended to use the recombinant protein formulated with a carrier protein such as BSA. Carrier proteins enhance stability, shelf-life, and allow for more dilute storage, which is advantageous for preparing standard curves. Carrier-free formulations are available but are typically reserved for applications where BSA may interfere with detection.

• Calibration and Parallelism: ELISA kits calibrated against NS0-expressed recombinant human HGF demonstrate that standard curves generated with this protein are linear and parallel to those obtained with natural HGF, allowing for accurate quantification of both recombinant and endogenous HGF in biological samples.

• Reconstitution and Handling: If using a lyophilized standard, reconstitute according to the manufacturer’s instructions, typically in PBS containing at least 0.1% BSA or human serum albumin to maintain protein stability and prevent adsorption to plasticware.

• Validation: Ensure that your ELISA antibodies recognize both recombinant and natural HGF, as most commercial kits are validated for this cross-reactivity. Always verify that your assay’s dynamic range and sensitivity are compatible with the concentrations expected in your samples.

• Documentation: For quantitative work, document the lot number, concentration, and formulation of your recombinant HGF standard, as minor batch-to-batch variations can affect calibration.

Summary Table: Use of Recombinant Human HGF (NS0-expressed) as ELISA Standard

In summary: Recombinant Human HGF (NS0 cell expressed) is suitable and commonly used as a standard for ELISA quantification, provided you follow best practices for formulation, reconstitution, and assay validation.

Applications of Recombinant Human HGF (NS0 Cell Expressed)

Recombinant Human HGF expressed in NS0 cells has been validated across a diverse range of scientific applications in published research, reflecting its broad biological activity.

Cell Culture and Bioassay Applications

The protein has demonstrated efficacy in cell culture systems and bioassay applications. Specifically, it has been validated for inducing IL-11 secretion by Saos-2 human osteosarcoma cells, with an ED₅₀ value equal to or less than 4.00 ng/mL.

Cellular Migration and Motility Studies

HGF has been extensively validated for studying cell migration across multiple cell types. The protein induces ERK phosphorylation and stimulates matrix metalloproteinase (MMP) production, thereby enhancing human myoblast migration on extracellular matrix (ECM) molecules such as laminin and fibronectin. Additionally, HGF promotes the motility of cardiac stem cells in damaged myocardium and serves as a chemoattractant for motor neurons.

Developmental and Regenerative Biology

The protein has been applied in research examining embryonic organ development, adult organ regeneration, and wound healing. HGF induces proliferation, motility, and morphological changes in thyrocytes while inhibiting TSH-stimulated iodine uptake. It also regulates the development of sensory, sympathetic, parasympathetic, and cortical neurons.

Metabolic and Immunological Functions

HGF has been validated for studying pancreatic beta cell function, supporting insulin production. The protein also supports neuronal survival and immune tolerance, making it valuable for immunological research applications.

Binding Activity Validation

The recombinant protein has been validated for binding activity assays, enabling researchers to study HGF receptor interactions and signaling mechanisms.

Human myoblasts, osteosarcoma cells, thyroid cells, cardiac stem cells, and various epithelial and endothelial cell types have all been successfully used with this preparation in published research.

To reconstitute and prepare Recombinant Human HGF (NS0 Cell Expressed) protein for cell culture experiments, dissolve the lyophilized protein at 50–100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA). This stabilizes the protein and prevents adsorption to surfaces.

Step-by-step protocol:

• Check the label or datasheet for the recommended reconstitution concentration, as it may vary by lot or formulation (commonly 50 μg/mL or 100 μg/mL).
• Add sterile PBS (phosphate-buffered saline) to the vial to reach the desired concentration.
• Include at least 0.1% BSA (or human serum albumin) in the PBS to stabilize the protein and minimize loss due to adsorption.
• Gently mix by pipetting or swirling; avoid vigorous vortexing to prevent denaturation.
• Allow the solution to sit at room temperature for 10–15 minutes to ensure complete dissolution.
• Aliquot the reconstituted protein into single-use volumes to avoid repeated freeze-thaw cycles, which can degrade activity.
• Store aliquots at –20°C to –70°C in a manual defrost freezer for long-term storage.

Additional notes:

• If the product is supplied without carrier protein (BSA), it is especially important to add BSA during reconstitution to prevent loss of activity.
• For cell culture experiments, dilute the reconstituted stock to the desired working concentration in your cell culture medium immediately before use.
• Avoid repeated freeze-thaw cycles, as this can reduce protein activity.
• Always use sterile technique to prevent contamination.

Typical working concentrations for cell culture applications range from 1–100 ng/mL, depending on the cell type and experimental design. Bioactivity can be confirmed by measuring induction of IL-11 secretion in responsive cell lines, with an ED₅₀ of ≤4 ng/mL reported for this protein.

Summary Table:

This protocol ensures optimal stability and bioactivity of recombinant HGF for cell culture experiments.