Recombinant Human NRG1-β1

Recombinant Human NRG1-β1 is widely used in research due to its critical roles in cell signaling, neurobiology, cardiovascular biology, and oncology. It is particularly valuable for studies involving neural development, nerve regeneration, cardiac function, and cancer cell biology.

Key scientific applications and rationale:

• Neural Development and Regeneration: NRG1-β1 promotes the growth, differentiation, and survival of neural cells, including neural stem cells and Schwann cells. It is essential for studying neural stem cell culture, nerve repair, and neuroprotection, as it can enhance functional nerve regeneration and reduce neuroinflammation.

• Neuroprotection and Disease Modeling: Exogenous NRG1-β1 protects neurons from oxidative stress and amyloid-beta-induced damage, making it a valuable tool in neurodegenerative disease models such as Alzheimer's disease. It modulates ErbB receptor signaling and downstream pathways (Akt, Erk1/2), which are implicated in neuronal survival and plasticity.

• Cardiovascular Research: Recombinant NRG1-β1 has demonstrated therapeutic potential in models of heart failure and myocardial injury. It regulates angiogenesis, blood pressure, and cardiac muscle responses, and exerts cytoprotective effects on cardiac and endothelial cells.

• Cancer Biology: NRG1-β1 stimulates proliferation in certain cancer cell lines (e.g., MCF-7 breast cancer cells) and is involved in tumorigenesis and metastasis. It is used to investigate ErbB receptor-mediated signaling in cancer progression and drug response.

• Cell Signaling Studies: NRG1-β1 is a ligand for ErbB3 and ErbB4 receptor tyrosine kinases, triggering heterodimerization with ErbB2 and activating downstream signaling cascades. This makes it a powerful tool for dissecting receptor biology and intracellular signaling mechanisms.

Best practices for use:

• Select appropriate concentrations based on cell type and experimental goals (commonly in the nanomolar range for in vitro studies).
• Use in defined media for neural or cardiac cell cultures to study differentiation, survival, or regeneration.
• Employ in disease models (e.g., neurodegeneration, heart failure, cancer) to assess therapeutic or mechanistic effects.

Summary of advantages:

• Enables precise modulation of ErbB signaling pathways.
• Facilitates mechanistic studies in neurobiology, cardiology, and oncology.
• Supports translational research for regenerative medicine and disease therapy.

In conclusion, Recombinant Human NRG1-β1 is a versatile and scientifically validated reagent for research applications requiring modulation of cell growth, differentiation, survival, and signaling, especially in neural, cardiac, and cancer biology contexts.

You can use recombinant human NRG1-β1 as a standard for quantification or calibration in your ELISA assays, provided it is of high purity and its concentration is accurately determined. This is a common practice in ELISA development and quantification workflows.

Key considerations and best practices:

• Purity and Quantification: The recombinant protein should be highly purified, and its concentration should be accurately measured, ideally by methods such as HPLC or absorbance at 280 nm.
• Standard Curve Preparation: Prepare a standard curve using serial dilutions of the recombinant NRG1-β1 in the same buffer or matrix as your samples to ensure accurate quantification.
• Immunoreactivity: The recombinant standard must contain the same epitope(s) recognized by the antibodies used in your ELISA. Most commercial ELISA kits for NRG1-β1 use recombinant human NRG1-β1 as the standard, confirming its suitability for this purpose.
• Lot-to-Lot Variability: Be aware that different lots of recombinant protein may have slight differences in immunoreactivity or mass determination, which can affect quantification. It is recommended to assign the standard curve concentration based on ELISA measurement rather than relying solely on the mass stated on the vial.
• Formulation: If your recombinant NRG1-β1 is supplied with carrier proteins (e.g., BSA), ensure this matches the matrix of your samples or use carrier-free protein if required for your assay.

Limitations:

• If your ELISA is designed to detect a specific isoform or domain, confirm that your recombinant standard matches the target sequence and structure.
• For absolute quantification, the standard should be as similar as possible to the endogenous protein in your samples in terms of post-translational modifications and folding.

Summary Table:

In summary, recombinant human NRG1-β1 is suitable as an ELISA standard if these criteria are met, and this approach is widely used in both commercial kits and custom assays.

Recombinant Human NRG1-β1 has been validated in published research for a diverse range of applications, primarily in cell-based bioassays, stimulation protocols, in vivo studies, and disease models, especially in neuroscience, cardiology, and oncology.

Key validated applications include:

• Cell-based bioassays: Used to stimulate proliferation, migration, and differentiation in various cell types, including cancer cell lines (e.g., MCF-7 breast cancer cells), neuronal progenitors, and glial cells.
• Cell culture stimulation: Applied to induce specific signaling pathways, such as ErbB receptor activation, and to study cellular responses in astrocytes, hepatocytes, and other primary cultures.
• In vivo studies: Administered in animal models (mouse, rat, monkey) to investigate effects on learning and memory, neuroinflammation, synaptic plasticity, and cardiac function.
• Cardiac regeneration and protection: Validated for stimulating cardiac regeneration in neonatal mice and improving cardiac function and survival in models of heart failure and ischemia/reperfusion injury.
• Disease modeling and therapeutic research: Used in preclinical and clinical studies for chronic systolic heart failure, schizophrenia, and cancer (breast, colorectal, head and neck squamous cell carcinoma).
• Neurobiology: Studied for its role in synaptic pruning, regulation of long-term potentiation (LTP), inhibition of neuroinflammatory responses, and modulation of myelination genes in glial cells.
• Tumor biology: Investigated for effects on tumor growth, migration, and resistance mechanisms in cancer models, including schwannomas and breast cancer.
• Biomarker research: Circulating NRG1-β1 levels have been explored as biomarkers for heart failure severity and risk stratification in clinical settings.

Representative published protocols and models:

• Subcutaneous administration in animal models for cardiac and neurological studies.
• Stimulation of cell lines and primary cultures to assess proliferation, migration, and signaling pathway activation.
• In vivo behavioral and physiological assays to evaluate learning, memory, and disease progression.

Summary Table: Applications of Recombinant Human NRG1-β1

In summary, recombinant human NRG1-β1 is a well-validated research tool for investigating cellular signaling, disease mechanisms, and therapeutic interventions in cardiovascular, neurological, and oncological contexts.

To reconstitute and prepare Recombinant Human NRG1-β1 protein for cell culture experiments, follow these steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Reconstitute the protein in sterile water or sterile PBS to a concentration of 0.1 mg/mL (100 μg/mL). If the product datasheet specifies, use PBS with at least 0.1% BSA to enhance stability and prevent adsorption to plastic surfaces.
• Gently pipette the solution down the side of the vial to dissolve the protein. Do not vortex; instead, gently swirl or invert the vial to avoid foaming and protein denaturation.
• Allow the protein to fully dissolve at room temperature for 15–30 minutes with gentle agitation.
• Aliquot the reconstituted protein into working volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• For long-term storage, dilute aliquots in a buffer containing 0.1% BSA and store at –80°C. For short-term use (2–7 days), store at 4°C.

Preparation for cell culture:

• Before adding to cell cultures, further dilute the reconstituted stock to the desired working concentration using cell culture medium or PBS with 0.1% BSA. Typical working concentrations for bioassays range from 0.06–0.3 ng/mL for cell proliferation assays, but optimal concentrations should be determined empirically for your specific application.
• Always use sterile technique throughout the process to prevent contamination.

Summary of best practices:

• Use sterile water or PBS (with or without BSA, depending on the application and product formulation).
• Avoid vigorous mixing; use gentle pipetting or swirling.
• Aliquot and store at –80°C for long-term use; avoid freeze-thaw cycles.
• Dilute to working concentration in cell culture medium immediately before use.

These steps ensure the protein remains stable and biologically active for your cell culture experiments.