Recombinant Human HGF R

Recombinant Human HGF R (hepatocyte growth factor receptor, also known as c-Met) offers several compelling advantages for research applications, particularly in studies involving angiogenesis, cell signaling, and tissue regeneration.

Key Biological Functions

The HGF R is a receptor tyrosine kinase that plays critical roles in multiple cellular processes. Activation of the c-Met signal transduction pathway triggers diverse cellular responses including cell motility, scattering, proliferation, survival, and angiogenesis. This makes it an essential tool for investigating vascular development and tissue repair mechanisms.

Advantages for Research

Superior Quality and Consistency

Recombinant proteins offer significant advantages over native equivalents, including superior bioactivity, batch-to-batch consistency, and purity. This consistency is particularly important for ensuring experimental reproducibility, which is a major challenge in modern research. When working with complex systems like organoids or tissue cultures, reliable reagents become essential for distinguishing genuine biological effects from artifacts caused by reagent variability.

Functional Validation

The biological activity of recombinant HGF R can be directly assessed through its ability to bind human HGF, allowing you to verify protein functionality before use in experiments. This quality control is critical for obtaining meaningful results in downstream applications.

Research Applications

Recombinant HGF R is particularly valuable for investigating therapeutic angiogenesis and vascular regeneration. Studies have demonstrated that HGF promotes collateral vessel development and can reduce symptoms of critical limb ischemia. Additionally, HGF in combination with other factors has shown promise in liver regeneration and stem cell induction, making recombinant HGF R useful for studying hepatic repair mechanisms and stem cell biology.

The receptor's role in multiple signaling pathways also makes it suitable for broader applications in inflammatory disease research and tissue engineering studies.

Yes, recombinant human HGF (Hepatocyte Growth Factor) can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for this purpose. Recombinant HGF is commonly used as a calibrator in commercial ELISA kits designed to quantify HGF in biological samples.

Key considerations and supporting details:

• Parallelism and Calibration: ELISA kits for human HGF are typically calibrated against highly purified recombinant human HGF. Validation studies show that standard curves generated with recombinant HGF are parallel to those obtained with natural HGF, allowing for accurate quantification of both forms. This parallelism is essential for reliable calibration.

• Standard Preparation: The recombinant HGF standard should be prepared according to the ELISA kit instructions, using the recommended diluent and concentration range. Most kits provide a standard curve covering a dynamic range suitable for quantifying HGF in serum, plasma, and cell culture supernatants.

• Assay Validation: It is important to confirm that your recombinant HGF preparation is compatible with the antibodies used in your ELISA. Commercial kits specify that their standards are recombinant HGF and demonstrate specificity for both recombinant and natural HGF. If using a custom or non-kit ELISA, ensure that your recombinant HGF is recognized by the capture and detection antibodies.

• Carrier Proteins: Some recombinant HGF preparations contain carrier proteins (e.g., BSA) to stabilize the protein. For ELISA calibration, carrier-free formulations are preferred unless the presence of carrier does not interfere with assay performance.

• Research Use Only: Most ELISA kits and recombinant standards are labeled for research use only and are not validated for diagnostic applications.

Best Practices:

• Use recombinant HGF from a reliable source with documented purity and activity.
• Validate the standard curve in your specific ELISA system, confirming linearity, accuracy, and parallelism with sample curves.
• Store and handle recombinant HGF according to manufacturer recommendations to maintain stability and activity.

Summary Table: Recombinant HGF as ELISA Standard

In conclusion, recombinant human HGF is suitable as a standard for ELISA quantification, provided assay validation confirms parallelism and specificity with your assay system.

Recombinant Human HGF R (Hepatocyte Growth Factor Receptor, c-Met) has been validated for several key applications in published research, primarily focused on its role in cell signaling, cancer biology, and regenerative medicine.

Validated Applications:

• Bioassays: Used to measure biological activity, such as binding to human HGF and activation of downstream signaling pathways.
• Cell Proliferation and Migration: Frequently employed to study cell proliferation, motility, and scattering, especially in cancer and angiogenesis models.
• Cell Culture: Utilized as a supplement in cell culture systems to stimulate cellular responses, including proliferation and survival.
• Immunoassay Standard: Used as a standard in ELISA and other immunoassays to quantify HGF or c-Met activity.
• In Vivo Studies: Applied in animal models to investigate therapeutic effects, such as neuroprotection, tissue regeneration, and antifibrotic activity.
• Activation and Control: Used to activate c-Met signaling in experimental systems and as a control protein in various assays.

Supporting Details:

• Cancer Research: Recombinant HGF R is widely used to study tumor growth, metastasis, and resistance to targeted therapies, given its central role in the c-Met pathway.
• Regenerative Medicine: Validated in studies on neural cell survival, axonal regeneration, and tissue repair, including clinical trials for spinal cord injury and vocal fold scarring.
• Angiogenesis: Used to investigate mechanisms of blood vessel formation and endothelial cell function.
• Fibrosis and Tissue Remodeling: Studied for its antifibrotic effects in models of liver, vocal fold, and other tissue injuries.

Experimental Protocols:

• Recombinant Human HGF R is typically used in concentrations ranging from sub-nanogram to microgram per milliliter, depending on the assay and cell type.
• Biological activity is confirmed by its ability to bind HGF and induce expected cellular responses, such as proliferation or migration.

Summary Table:

Note: The above applications are supported by published research and product validation data. Specific protocols and concentrations should be optimized for each experimental system.

To reconstitute and prepare Recombinant Human HGF R (c-MET) protein for cell culture experiments, follow these best-practice steps:

• Briefly centrifuge the vial before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute the protein in sterile 1× PBS (phosphate-buffered saline), pH 7.2–7.4. If the product datasheet specifies, use PBS without calcium or magnesium and with a carrier protein such as 0.1% endotoxin-free human or bovine serum albumin (HSA/BSA) to stabilize the protein and prevent adsorption to surfaces.
• Typical reconstitution concentration is 0.1–1.0 mg/mL; a common working concentration is 0.2 mg/mL. For example, to reconstitute 100 µg of protein at 0.2 mg/mL, add 0.5 mL of buffer.
• Gently mix by swirling or tapping the vial. Avoid vigorous shaking or vortexing, which can denature the protein or cause foaming.
• Allow the protein to fully dissolve at room temperature for 15–30 minutes with gentle agitation.
• Aliquot the reconstituted protein into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage after reconstitution:
  • Short-term (up to 1 week): 2–8 °C.
  • Long-term (up to 6 months): –20 °C to –80 °C.
  • Avoid multiple freeze-thaw cycles.

Summary Table: Reconstitution and Storage

Additional Notes:

• Always consult the product-specific Certificate of Analysis (CoA) or datasheet for any unique requirements.
• Use only sterile, endotoxin-free reagents and plasticware for cell culture applications.
• If using for sensitive cell types or stem cells, ensure all components are xeno-free and endotoxin levels are below 1 EU/µg.

These steps will help ensure the biological activity and stability of Recombinant Human HGF R protein for your cell culture experiments.