Recombinant Mouse NGF R

Recombinant Mouse Nerve Growth Factor (NGF), often referred to as Recombinant Mouse β-NGF, is a widely used tool in neuroscience and regenerative research due to its well-characterized biological activity and relevance to neuronal survival, differentiation, and repair. Here are several reasons why you might choose to use Recombinant Mouse NGF in your research applications:

1. Species-Specific Biological Activity

Recombinant Mouse NGF is specifically designed to interact with mouse TrkA and p75 receptors, making it ideal for studies involving mouse models. Using species-matched NGF ensures optimal receptor binding and signaling, which is critical for accurate in vitro and in vivo experiments.

2. Neuronal Survival and Differentiation

NGF is essential for the survival, maintenance, and differentiation of sympathetic and sensory neurons. Recombinant Mouse NGF is routinely used to:

• Promote neurite outgrowth in primary neuronal cultures.
• Support the survival of neurons in culture.
• Induce differentiation of cell lines such as PC12 cells, a classic model for studying neuronal differentiation.

3. Neuroprotection and Regeneration Studies

Mouse NGF has been shown to prevent neuronal degeneration in various animal models and is a therapeutic target in studies of nerve regeneration. It can be used to:

• Investigate mechanisms of neuroprotection in models of neurodegeneration.
• Assess the potential of NGF in promoting recovery after nerve injury.

4. Standardization and Reproducibility

Recombinant Mouse NGF is produced under controlled conditions, ensuring batch-to-batch consistency, high purity, and the absence of contaminants (such as carrier proteins, if using carrier-free versions). This standardization is crucial for reproducible experimental results.

5. Compatibility with Common Assays

Recombinant Mouse NGF is compatible with a wide range of assays, including:

• Cell culture and differentiation assays.
• In vivo animal studies (e.g., intracerebral, intraperitoneal, or topical administration).
• Immunohistochemistry and biochemical analyses of neuronal survival and signaling pathways.

6. Well-Established Protocols and Literature Support

There is a large body of published research using Recombinant Mouse NGF, providing validated protocols and reference data for experimental design and interpretation.

7. Research Applications

Recombinant Mouse NGF is suitable for studies in:

• Neurodevelopment.
• Neurodegenerative diseases (e.g., Alzheimer’s, Parkinson’s).
• Peripheral neuropathies.
• Pain research.
• Retinal and optic nerve studies.

In summary, Recombinant Mouse NGF is a reliable, biologically active reagent that supports a broad range of neuroscience research applications, particularly when working with mouse models or cell lines. Its use ensures species-specific activity, reproducibility, and compatibility with established experimental protocols.

You can use recombinant Mouse NGF R as a standard for quantification or calibration in ELISA assays, provided that the recombinant protein is well-characterized, pure, and matches the analyte detected by your assay. However, several technical considerations must be addressed to ensure accurate quantification:

• Protein Identity and Form: The standard must be the same molecular form (e.g., mature NGF, proNGF, or NGF receptor) as the analyte your ELISA is designed to detect. For example, if your ELISA is specific for mature NGF, using recombinant proNGF or NGF receptor (NGF R/TNFRSF16) as a standard will not yield accurate results due to differences in antibody recognition and epitope specificity.

• Validation: The recombinant standard should be validated for use in your specific ELISA format. Some kits are optimized for natural proteins purified from tissue, while others are validated for recombinant proteins. If your ELISA kit documentation or technical manual specifies compatibility with recombinant Mouse NGF R, it can be used as a standard.

• Purity and Quantification: The recombinant protein should be highly purified, with its concentration accurately determined (e.g., by BCA assay or absorbance at 280 nm). Impurities or incorrect quantification will affect the standard curve and assay accuracy.

• Expression System: Recombinant proteins expressed in different systems (e.g., E. coli, mammalian cells) may have different post-translational modifications, affecting antibody binding and assay performance. Ensure the recombinant Mouse NGF R matches the form recognized by your ELISA antibodies.

• Calibration Curve Preparation: Prepare serial dilutions of the recombinant standard in the same buffer as your samples to minimize matrix effects. Follow best practices for standard curve generation, including running standards in duplicate or triplicate.

Key technical points:

• If your ELISA is designed for NGF R/TNFRSF16 detection, recombinant Mouse NGF R is appropriate as a standard.
• If your ELISA is for mature NGF or proNGF, use the corresponding recombinant protein as the standard, not NGF R.
• Always consult your ELISA kit documentation for recommended standards and validation data.

In summary, recombinant Mouse NGF R can be used as a standard for ELISA quantification only if it matches the analyte and is validated for your assay format. Otherwise, use the specific recombinant protein (mature NGF, proNGF) that corresponds to your assay target.

Recombinant Mouse NGF R (Nerve Growth Factor Receptor, also known as p75NTR or NGFR) has been validated in published research primarily for applications involving ligand binding studies, functional antagonism of NGF, and investigations of neurotrophin signaling.

Key validated applications in published research include:

• Surface Plasmon Resonance (SPR): Used to characterize the high-affinity binding of recombinant NGF R/Fc chimera to NGF, confirming its utility in quantitative ligand-receptor interaction studies.
• NGF Antagonism in Cell-Based Assays: The recombinant NGF R/Fc chimera acts as a potent NGF antagonist, blocking NGF-mediated signaling in neuronal and non-neuronal cells, and is used to dissect NGF-dependent pathways in vitro.
• Studies of Cell Migration, Gene Expression, and Apoptosis: NGF R has been employed to investigate its regulatory roles in cell migration, gene expression, and apoptosis, both as a co-receptor with TrkA and as an independent signaling molecule.
• Developmental and Adult Expression Analysis: Recombinant NGF R has been used as a probe to study the expression and function of p75NTR in various tissues during development and in adult organisms, including neuronal and non-neuronal cell types.
• Neurotrophin Signaling Modulation: Used in studies examining the modulation of neurotrophin signaling, including the association of NGF R with Trk receptors and sortilin to enhance neurotrophin transport and signaling.

Additional context:

• Recombinant NGF R is often used in its Fc-chimera form to facilitate purification, detection, and functional studies.
• It is validated for use in both cell and tissue culture systems, and as a standard in ELISA assays for quantifying NGF or NGF R interactions.
• Naturally occurring truncated forms of NGF R (extracellular domain only) have been detected in biological fluids, supporting its physiological relevance as a soluble NGF antagonist.

Summary Table:

If you require details on a specific experimental protocol or a particular disease model, please specify for more targeted information.

To reconstitute and prepare Recombinant Mouse NGF R (p75NTR/NGFR) protein for cell culture experiments, follow these best-practice steps based on current protocols for recombinant neurotrophin receptors:

1. Bring the vial to room temperature before opening to prevent condensation from forming inside the vial.

2. Centrifuge the vial briefly to ensure all lyophilized protein is collected at the bottom.

3. Reconstitution buffer selection:

   • If the product is lyophilized, use sterile PBS or sterile distilled water as the solvent, unless the datasheet specifies otherwise.
   • For increased stability and to prevent adsorption to plastic, add a carrier protein such as 0.1–1% BSA (bovine serum albumin) or HSA (human serum albumin) to the buffer.

4. Reconstitution concentration:

   • Typical stock concentrations are 0.1–1.0 mg/mL (100–1000 μg/mL).
   • For example, to prepare a 1 mg/mL stock, add 100 μL of buffer to 100 μg of protein.
   • For NGFR-Fc chimeras, stock solutions are often prepared at 50–100 μg/mL in the chosen buffer with carrier protein.

5. Dissolve the protein gently:

   • Do not vortex. Gently pipette up and down or swirl to dissolve. Allow the solution to sit at room temperature for 15–30 minutes with gentle agitation if needed.
   • Avoid vigorous shaking or foaming, which can denature the protein.

6. Aliquot and storage:

   • Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
   • Store aliquots at –20°C or colder (preferably –70°C for long-term storage).
   • For short-term use (up to 1 week), aliquots can be kept at 2–8°C.
   • Avoid repeated freeze/thaw cycles, as this can reduce activity.

7. Working solution preparation:

   • Before adding to cell culture, dilute the stock solution to the desired working concentration using cell culture medium or buffer containing 0.1% BSA/HSA to maintain stability and bioactivity.

Key technical notes:

• Always consult the specific product datasheet for any unique requirements, as formulation and recommended buffers may vary by manufacturer.
• If the protein is provided in solution (not lyophilized), it is typically already in PBS and may only require dilution and aliquoting.
• For functional assays, confirm the absence of cytotoxic preservatives and ensure endotoxin levels are suitable for cell culture (<1 EU/μg is typical).

Summary protocol example for lyophilized Recombinant Mouse NGF R:

1. Briefly centrifuge the vial.
2. Add sterile PBS with 0.1% BSA to achieve 0.1–1.0 mg/mL.
3. Gently mix and allow to dissolve at room temperature.
4. Aliquot and store at –20°C or colder.
5. Dilute to working concentration in cell culture medium with 0.1% BSA before use.

This approach ensures protein stability, preserves bioactivity, and minimizes loss due to adsorption or degradation.