Recombinant Human NRG1-α

Recombinant Human NRG1-α is used in research applications primarily for its roles in cardiac repair, neuroprotection, and cellular signaling, making it a valuable tool for studies in cardiovascular, neurological, and developmental biology.

Key scientific applications and rationale:

• Cardiac Function and Repair: NRG1-α is a member of the neuregulin family, which interacts with ErbB receptors to regulate cardiomyocyte survival, proliferation, and adaptation to injury. Recombinant human NRG1 has been shown to improve cardiac output, reduce apoptosis, and promote regeneration in both animal models and human studies of heart failure and myocardial injury. Its use enables mechanistic studies of cardiac signaling pathways and therapeutic evaluation for heart disease.

• Neuroprotection and Neural Development: NRG1-α is involved in neuronal survival, axon regeneration, and protection against oxidative stress. It has demonstrated efficacy in reducing neuronal death, neuroinflammation, and oxidative damage in models of stroke and neurodegenerative diseases. This makes it suitable for research into neuroprotective strategies and nervous system development.

• Cellular Signaling and Developmental Biology: NRG1-α regulates key processes in embryonic development, including nervous system, heart, and mammary gland formation. It is also implicated in cell adhesion, migration, and differentiation, allowing researchers to dissect signaling pathways and gene regulation in various cell types.

• Disease Modeling and Therapeutic Screening: Recombinant NRG1-α provides a controlled, reproducible reagent for in vitro and in vivo studies, enabling the modeling of disease states (e.g., heart failure, stroke, neurodegeneration) and the screening of potential therapeutic interventions targeting NRG1/ErbB pathways.

Best practices for use:

• Employ recombinant NRG1-α in dose-response and time-course experiments to elucidate its effects on cell survival, proliferation, and signaling.
• Use in combination with pathway inhibitors or genetic models to dissect downstream signaling mechanisms.
• Validate effects in relevant primary cells, organoids, or animal models for translational relevance.

Summary of scientific value:
Using recombinant human NRG1-α in research enables precise investigation of its biological functions, therapeutic potential, and underlying mechanisms in cardiovascular, neurological, and developmental contexts, supporting both basic science and translational studies.

Yes, recombinant human NRG1-α can be used as a standard for quantification or calibration in ELISA assays, provided that the ELISA kit is designed to detect the NRG1-α isoform and the recombinant protein is compatible with the assay system.

Key Points:

• Compatibility: The recombinant human NRG1-α protein should match the isoform and epitope recognized by the antibodies in your ELISA kit. For example, if your kit detects the EGF-like domain (common to all NRG1 isoforms), most recombinant NRG1-α proteins will be suitable.
• Purity and Activity: Ensure the recombinant protein is highly purified and, if possible, biologically active. Some suppliers provide proteins specifically validated for use as ELISA standards.
• Carrier Protein: If the recombinant protein is supplied with a carrier (e.g., BSA), make sure this does not interfere with your ELISA. Carrier-free versions are preferred for ELISA standards.
• Validation: Always validate the standard curve using the recombinant protein in your specific ELISA protocol to confirm linearity and accuracy.

Example Suppliers and Products:

• R&D Systems: Recombinant Human NRG1/HRG1 Protein, CF (5898-NR) – suitable for use as an ELISA standard.
• Bio-Techne: Recombinant Human NRG1-alpha/HRG1-alpha EGF Domain Protein – suitable for ELISA standards.
• Novus Biologicals: Human NRG1 Recombinant Protein – fully biologically active and suitable for ELISA standards.

Protocol Considerations:

• Reconstitute the lyophilized recombinant protein according to the manufacturer's instructions.
• Prepare a series of dilutions to generate a standard curve.
• Run the standard curve alongside your samples to ensure accurate quantification.

By following these guidelines, you can effectively use recombinant human NRG1-α as a standard in your ELISA assays.

Recombinant Human NRG1-α has been validated for several important applications across both basic research and therapeutic development:

Cell-Based Bioassays

The primary validated application is cell proliferation assays, particularly using MCF-7 human breast adenocarcinoma cells in serum-free conditions. This assay measures the biological activity of the protein by quantifying its ability to stimulate cell growth, with established ED50 values documented in the literature.

Developmental and Neurobiological Research

NRG1-α has been extensively validated in studies examining its roles in nervous system development. Research has demonstrated its involvement in regulating neurotransmitter receptor expression in neurons and at the neuromuscular junction, as well as promoting Schwann cell differentiation from neural crest stem cells. The protein has also been validated for studying oligodendroglial lineage establishment.

Cardiac and Muscle Biology

Published research has validated NRG1-α for investigating its effects on skeletal muscle denervation and reinnervation through modulation of the Neuregulin 1/ErbB system. Additionally, the protein has been used to study myocardial development, including trabeculation of the developing heart and cardiomyocyte function.

In Vivo Therapeutic Studies

Recombinant NRG1 preparations have been validated in animal models for neuroprotection and cardioprotection. Studies have demonstrated its efficacy in attenuating mortality associated with experimental cerebral malaria and providing neuroprotection during ischemic stroke through reduction of inflammatory responses. Clinical trials have also evaluated recombinant NRG1 preparations for heart regeneration and cardiovascular indications.

Receptor Signaling Studies

The protein's validated application includes studying ErbB receptor signaling, as all NRG1 isoforms contain an EGF-like domain required for direct binding to ErbB3 or ErbB4 receptor tyrosine kinases.

To reconstitute and prepare Recombinant Human NRG1-α protein for cell culture experiments, dissolve the lyophilized protein in sterile water or sterile PBS at a concentration of 100 μg/mL or higher. After reconstitution, further dilute the stock solution in cell culture medium or buffer as needed for your experiment.

Step-by-step protocol:

1. Centrifuge the vial briefly to ensure all lyophilized powder is at the bottom before opening.
2. Add sterile water or PBS:
   • For most preparations, use sterile deionized water or sterile PBS.
   • If recommended by the product datasheet, include 0.1% BSA or HSA as a carrier protein to stabilize the solution, especially for long-term storage or low concentration applications.
3. Reconstitution concentration:
   • Prepare a stock solution at ≥100 μg/mL (e.g., dissolve 100 μg protein in 1 mL solvent).
   • For some protocols, concentrations up to 1 mg/mL are acceptable.
4. Mix gently by pipetting up and down or gentle vortexing until fully dissolved. Avoid vigorous agitation to prevent protein denaturation.
5. Aliquot the stock solution to avoid repeated freeze-thaw cycles.
6. Storage:
   • Short-term: Store at 4°C for up to 1 week.
   • Long-term: Store aliquots at –20°C to –80°C.
   • Avoid repeated freeze-thaw cycles to maintain protein activity.
7. Dilution for cell culture:
   • Dilute the stock solution into your cell culture medium to the desired working concentration immediately before use.
   • If using serum-free medium, consider adding carrier protein (e.g., 0.1% BSA) to prevent adsorption to plasticware.

Additional notes:

• Always consult the specific product datasheet for any unique requirements regarding buffer composition, pH, or additives.
• For bioassays, typical working concentrations range from 0.1–100 ng/mL, depending on cell type and experimental design.
• If the protein is carrier-free, extra care should be taken to avoid loss due to adsorption; use low-binding tubes and add carrier protein if compatible with your assay.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of Recombinant Human NRG1-α for cell culture experiments.