Recombinant Human NGF R

Recombinant Human Nerve Growth Factor (rhNGF) is a powerful research tool with broad applications in neuroscience, regenerative medicine, and disease modeling. Here are several compelling reasons to use rhNGF in your research:

1. Neuroprotection and Neuronal Survival

rhNGF is a well-characterized neurotrophic factor that promotes the survival, growth, and differentiation of neurons, particularly sensory and sympathetic neurons. In research models of neurodegeneration (such as glaucoma, optic neuropathy, and Alzheimer’s disease), rhNGF has been shown to:

• Reduce neuronal apoptosis (programmed cell death)
• Enhance survival of retinal ganglion cells (RGCs) and their axons
• Inhibit astrocyte activation, which can contribute to neuroinflammation and secondary degeneration

2. Relevance to Disease Models

rhNGF is widely used in preclinical studies to model and investigate:

• Optic neuropathies (e.g., glaucoma, traumatic optic nerve injury)
• Peripheral neuropathies
• Neurodegenerative diseases (e.g., Alzheimer’s disease)
• Corneal and cutaneous wound healing

Its effects are reproducible and well-documented in both rodent and primate models, making it a reliable tool for translational research.

3. Functional and Structural Recovery

Studies have demonstrated that rhNGF not only improves neuronal survival but also supports functional recovery, such as:

• Restoration of visual function in animal models and some clinical cases
• Enhanced axonal regeneration and synaptic connectivity
• Improved wound healing in skin and cornea

4. High Biological Activity and Purity

Modern recombinant production methods yield rhNGF with high purity and biological activity, comparable to native human NGF. This ensures consistent and reproducible results in experimental settings.

5. Versatile Delivery Methods

rhNGF can be administered via multiple routes, including:

• Topical (e.g., eye drops for retinal studies)
• Intravitreal or intraneural injection
• Systemic delivery

This flexibility allows researchers to tailor delivery methods to their specific experimental needs.

6. Translational Potential

rhNGF is already being evaluated in clinical trials for various indications, including:

• Glaucoma
• Peripheral neuropathy
• Corneal ulcers
• Pressure ulcers

Using rhNGF in your research aligns your work with ongoing clinical efforts and increases the translational relevance of your findings.

7. Well-Established Assays and Protocols

There are established protocols for assessing the effects of rhNGF, including:

• Immunohistochemistry (e.g., Brn3a staining for RGCs)
• Apoptosis assays (e.g., DARC imaging)
• Functional assessments (e.g., electroretinography, visual evoked potentials)

These tools facilitate robust and quantitative analysis of rhNGF’s effects.

In summary, recombinant Human NGF is a versatile, biologically active, and clinically relevant molecule that can significantly enhance the quality and impact of your research in neuroscience, neuroprotection, and regenerative medicine.

No, you cannot use recombinant human NGF R as a standard for NGF quantification in ELISA assays. NGF R (also known as CD271 or TNFRSF16) is the receptor for NGF, not the ligand itself, and these are distinct proteins that serve different functions in the assay system.

Key Distinctions

NGF versus NGF R: Nerve growth factor (NGF) is the protein ligand being measured in quantitative ELISA assays, while NGF R is the cell surface receptor that binds to NGF. They are structurally and functionally different molecules. ELISA kits designed to measure NGF require NGF standards, not NGF R standards, because the antibody pairs in the sandwich ELISA are specifically configured to capture and detect NGF molecules.

Appropriate Standards for NGF Quantification: For NGF ELISA assays, you should use recombinant human NGF standards that are specifically approved for this purpose. The WHO-approved recombinant human NGF standard, typically produced in mammalian expression systems, is the appropriate calibration reagent. The standard curve should be prepared using purified or recombinant protein that matches the target analyte—in this case, mature NGF or beta-NGF.

Important Considerations for NGF Standardization

ProNGF Interference: When establishing your standard curve, be aware that recombinant proNGF (the precursor form) can reciprocally interfere with NGF quantification in ELISA assays. Most commercial NGF ELISA kits preferentially detect mature NGF over proNGF, with cross-reactivity typically less than 0.1%, but this distinction is critical for accurate quantification.

Sample-Specific Requirements: If you are working with citrate-plasma samples, heterophilic antibody blockers may be necessary to prevent non-specific binding that could compromise your standard curve accuracy.

For your ELISA calibration, ensure you are using the correct recombinant NGF standard that matches your assay's specifications and target form of the protein.

Recombinant Human NGF (nerve growth factor) has been validated in published research for applications including neuroprotection, nerve regeneration, wound healing, and as a therapeutic agent in both in vitro and in vivo models, particularly for ocular and neurodegenerative diseases.

Key validated applications in published research:

• Neuroprotection and Neuroregeneration

  • Promotes survival and regeneration of neurons in models of neurodegenerative diseases, such as Alzheimer's disease and experimental allergic encephalomyelitis.
  • Demonstrated neuroprotective effects on retinal ganglion cells (RGCs) and axons in optic neuropathy models, reducing apoptosis and promoting axonal survival.
  • Used in studies of peripheral neuropathies, including diabetic polyneuropathy, where it improved clinical signs and symptoms in phase II and III clinical trials.

• Ophthalmic Applications

  • Validated for stimulation of corneal wound healing, improvement of corneal sensitivity, and regeneration of corneal nerves in both animal models and human clinical trials.
  • Used as a topical agent (eye drops) for treating neurotrophic keratitis, dry eye disease, and other ocular surface diseases, with demonstrated efficacy in improving nerve density and epithelial healing.
  • Incorporated into bioengineered corneal lenticules for sustained release and therapeutic effect in corneal repair.

• Wound Healing and Tissue Repair

  • Shown to promote healing of cutaneous wounds, including corneal and pressure ulcers, and to enhance mast cell activity and tissue regeneration in vivo.

• Bioassays and In Vitro Studies

  • Used in bioactivity assays to assess neuronal differentiation, neurite outgrowth, and cell survival in cultured cells.
  • Demonstrated ability to stimulate growth of corneal epithelial cells and maintain limbal epithelial stem cell potential in vitro.

• Clinical Trials and Therapeutic Use

  • Recombinant human NGF has been evaluated in multiple clinical trials for peripheral neuropathies, ocular diseases, and neurodegenerative conditions, with several studies reporting positive outcomes and FDA approval for specific indications such as neurotrophic keratitis.

Summary Table: Validated Applications of Recombinant Human NGF

Experimental techniques validated with recombinant human NGF include:

• Topical administration (eye drops)
• Subcutaneous or intramuscular injection
• Incorporation into biomaterials for sustained release
• Immunohistochemistry, confocal microscopy, and cell-based bioassays

These applications are supported by a robust body of preclinical and clinical research, with recombinant human NGF now established as a key tool in neuroscience, ophthalmology, and regenerative medicine.

To reconstitute and prepare Recombinant Human NGF R protein for cell culture experiments, first briefly centrifuge the vial to collect the lyophilized powder at the bottom. Use sterile technique throughout the process.

Reconstitution Protocol:

• Add sterile, distilled water or sterile PBS (pH 7.4) as the diluent, depending on the specific formulation and experimental requirements.
• For most applications, reconstitute to a final concentration of 0.1–1.0 mg/mL. For example, add 1 mL of diluent to 1 mg of protein for a 1 mg/mL solution.
• If recommended, include a carrier protein such as 0.1% human or bovine serum albumin (HSA/BSA) to stabilize the protein, especially for long-term storage or low concentration use.
• Do not vortex or mix vigorously; instead, gently swirl or tap the vial to dissolve the protein and avoid foaming.
• Allow the solution to sit at room temperature for several minutes to ensure complete dissolution.

Preparation for Cell Culture:

• After reconstitution, dilute the stock solution to the desired working concentration using sterile cell culture medium or buffer. Typical working concentrations for bioassays range from 0.2–2 ng/mL, but optimal dosing should be determined empirically for your cell type and assay.
• Filter-sterilize the final working solution if required for sensitive cell culture applications.

Storage Recommendations:

• Store the reconstituted protein at 2–8°C for up to 1 week.
• For longer-term storage, aliquot and freeze at –20°C to –70°C to avoid repeated freeze-thaw cycles.
• If carrier protein is included, stability may be enhanced for extended storage.

Additional Notes:

• Always consult the product-specific datasheet or Certificate of Analysis for any unique requirements regarding diluent, concentration, or additives.
• Avoid reconstituting to concentrations below 100 µg/mL unless specifically recommended, as lower concentrations may reduce stability.
• Use aseptic technique to prevent contamination.

This protocol ensures optimal solubility, stability, and biological activity of recombinant NGF R protein for cell culture experiments.