Recombinant Mouse Amphiregulin

Recombinant mouse amphiregulin (AR) is a valuable tool for research applications due to its well-characterized biological functions and versatility across multiple experimental contexts.

Biological Activity and Cellular Effects

Recombinant mouse amphiregulin demonstrates robust dose-dependent proliferative effects on multiple cell types. The protein stimulates proliferation of fibroblasts, keratinocytes, epithelial cells, astrocytes, and glial cells, making it suitable for studies examining growth factor signaling and cellular proliferation. The full-length 98 amino acid recombinant form exhibits superior receptor binding and biological activity compared to truncated variants, ensuring reliable and reproducible results in your experiments.

Tissue Repair and Regeneration

Beyond proliferation, recombinant amphiregulin plays a critical role in tissue repair and homeostasis. In vitro studies demonstrate that the protein significantly enhances wound healing in mesangial cells, tubular cells, and glomerular endothelial cells. The protein also facilitates angiogenesis, as evidenced by tube formation assays with endothelial cells. These properties make it particularly valuable for investigating mechanisms of epithelial regeneration, tissue integrity maintenance, and wound healing responses.

Diverse Research Applications

The protein is applicable across multiple experimental paradigms:

• Bioassays for measuring cellular responses and growth factor signaling
• In vivo assays using transgenic mouse models
• Organoid studies examining three-dimensional tissue development

Immunological and Pathological Relevance

Recombinant amphiregulin is instrumental for studying immune regulation and disease mechanisms. The protein is relevant to investigations of regulatory T cell function, inflammation resolution, tumor microenvironment dynamics, and tissue-specific pathologies including cancer, fibrosis, and infection-induced tissue damage. Its expression patterns in various carcinomas and inflammatory conditions make it essential for cancer biology and immunology research.

The combination of well-established biological activity, multiple cell type responsiveness, and applicability across diverse experimental systems makes recombinant mouse amphiregulin a robust choice for mechanistic studies of growth factor signaling, tissue regeneration, and immune regulation.

Yes, you can use recombinant mouse amphiregulin as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is suitable for this purpose and is compatible with your specific ELISA kit.

Key Points:

• Recombinant Protein as Standard: Recombinant mouse amphiregulin is commonly used as a standard in ELISA assays to generate a standard curve, which allows for the quantification of amphiregulin levels in your samples. This is especially true if the ELISA kit is designed to detect both natural and recombinant forms of the protein.

• Kit Compatibility: Most commercial ELISA kits for mouse amphiregulin (such as those from Invitrogen, Abcam, R&D Systems, and others) are validated to recognize both natural and recombinant amphiregulin. Always check the kit manual or datasheet to confirm compatibility.

• Purity and Form: For best results, use a highly purified recombinant amphiregulin, preferably with a known concentration and activity. Some suppliers offer recombinant proteins specifically labeled as "ELISA standards" or "for use as standards."

• Carrier Protein: If the recombinant amphiregulin is supplied with a carrier protein (such as BSA), this may affect the standard curve if your samples do not contain the same carrier. Carrier-free versions are preferred for standard curves.

• Calibration: When using recombinant protein as a standard, ensure that it is diluted in the same matrix as your samples (e.g., assay diluent, serum, or buffer) to minimize matrix effects.

Example Suppliers:

• Bio-Techne (R&D Systems): Offers recombinant mouse amphiregulin (e.g., 989-AR) that is calibrated for use in ELISA.
• BioLegend: Provides carrier-free recombinant mouse amphiregulin suitable for ELISA standards.
• Sigma-Aldrich: Also offers recombinant mouse amphiregulin for use in ELISA.

Recommendation:

Always refer to the manufacturer's instructions for both the ELISA kit and the recombinant protein to ensure compatibility and optimal performance. If in doubt, contact the technical support of the kit or protein supplier for guidance.

Recombinant Mouse Amphiregulin has been validated for a wide range of applications in published research, including:

• Bioassay: Used to assess biological activity, such as proliferation of fibroblasts, keratinocytes, and epithelial cells, as well as tissue repair and immune modulation in various cell types and organoids.
• In vivo assay: Employed in animal models to study tissue repair, immune suppression, lung regeneration, liver regeneration, and tumor growth.
• Functional Assay: Validated for evaluating the functional effects of amphiregulin, such as its role in cell proliferation, tissue homeostasis, and signaling pathways.
• Blocking/Neutralization Assay: Used to study the effects of inhibiting amphiregulin signaling, particularly in contexts like immune regulation and tissue repair.
• ELISA: Applied as a standard or in detection assays to quantify amphiregulin levels.
• Western Blot: Used for detection and analysis of amphiregulin protein expression.

These applications are supported by studies investigating amphiregulin's roles in tissue repair, immune regulation, cancer, and developmental biology.

To reconstitute and prepare Recombinant Mouse Amphiregulin protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer (commonly PBS or sterile distilled water) to a concentration suitable for your application, typically between 0.1–1.0 mg/mL. Avoid vigorous mixing and repeated freeze-thaw cycles to maintain protein integrity.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all contents are at the bottom.
• Reconstitution:
  • Add sterile PBS or sterile distilled water to achieve the desired concentration (e.g., 0.1–1.0 mg/mL).
  • For enhanced stability, you may add a carrier protein such as 0.1–1% BSA or HSA, especially at low working concentrations.
  • Gently mix by pipetting or slow inversion; do not vortex as this may destabilize the protein.
• Aliquot the solution to avoid repeated freeze-thaw cycles.
• Storage:
  • Store aliquots at -20°C or -80°C for long-term use.
  • For short-term use (up to one week), aliquots can be kept at 2–8°C.
• Working solution preparation:
  • Dilute the stock solution in cell culture medium or appropriate buffer immediately before use.
  • Typical working concentrations for cell stimulation range from 5–50 ng/mL, depending on cell type and experimental design.
• Handling precautions:
  • Avoid repeated freeze-thaw cycles.
  • Use freshly prepared working solutions for best activity.

Summary Table:

Additional notes:

• Always consult the specific product datasheet for recommended buffer and concentration, as formulations may vary.
• If the protein is supplied in a buffer containing stabilizers (e.g., trehalose, BSA), adjust your protocol accordingly.
• For cell culture, ensure all solutions are sterile and endotoxin levels are suitable for your application (<0.1 EU/μg is typical for sensitive assays).

This protocol ensures optimal solubility, stability, and biological activity of recombinant mouse Amphiregulin for cell culture experiments.