Recombinant Mouse EGF

Recombinant Mouse EGF is widely used in research because it is a potent, well-characterized growth factor that stimulates cell proliferation, differentiation, and survival in a variety of mouse-derived cell types and tissues. Its recombinant form ensures high purity, batch-to-batch consistency, and reliable biological activity, which are critical for reproducible experimental results.

Key reasons to use recombinant mouse EGF in research applications:

• Cell Proliferation and Differentiation: EGF robustly induces proliferation and differentiation of epithelial cells, fibroblasts, and stem cells, making it essential for studies involving tissue regeneration, wound healing, and organoid culture.
• Reproducibility and Consistency: Recombinant production provides high bioactivity and minimizes variability between batches, which is crucial for reproducible results, especially in sensitive applications like organoid culture and stem cell research.
• Defined Activity: Recombinant EGF is free from animal-derived contaminants and has a defined, quantifiable activity, allowing precise control over experimental conditions.
• Versatility: It is used in a wide range of applications, including:
  • Maintenance and expansion of stem cells and organoids.
  • Bioassays for cell signaling, proliferation, and survival studies.
  • Studies of epithelial development, angiogenesis, and tissue repair.
  • Investigation of disease models, such as cancer, neurogenesis, and metabolic disorders.
• Species Specificity: Using mouse EGF is important for mouse cell and tissue models to ensure optimal receptor binding and physiological relevance, as cross-species differences can affect activity.

Additional context:

• EGF acts by binding to the EGF receptor (EGFR), triggering receptor dimerization and autophosphorylation, which activates downstream signaling pathways involved in cell growth and survival.
• Recombinant EGF is also used in dermatological research for wound healing, skin regeneration, and anti-aging studies due to its effects on fibroblast activity and collagen production.

In summary, recombinant mouse EGF is a critical tool for mouse cell biology, tissue engineering, and disease modeling, offering high reliability, defined activity, and broad applicability across research fields.

Yes, recombinant mouse EGF can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay requirements. Recombinant EGF is commonly used as the standard in commercial mouse EGF ELISA kits, where its concentration is used to generate a standard curve for quantifying EGF in biological samples.

Key considerations and best practices:

• Validation: The recombinant mouse EGF standard should be validated for use in your specific ELISA system. Commercial kits typically use recombinant EGF expressed in E. coli or other systems, and their standard curves have been shown to be parallel to those obtained with natural mouse EGF, indicating suitability for quantification.
• Purity and Activity: Ensure the recombinant EGF is of high purity and retains biological activity. Most commercial standards are >98% pure and tested for activity.
• Matrix Effects: When quantifying EGF in complex biological samples (e.g., serum, plasma, tissue homogenates), confirm that the recombinant standard behaves similarly to native EGF in your assay matrix. Recovery and parallelism studies in commercial kits demonstrate that recombinant EGF can be reliably used for calibration across various sample types.
• Storage and Handling: Follow recommended storage conditions (typically -20 °C or -80 °C for lyophilized standards) and avoid repeated freeze/thaw cycles to maintain protein integrity.
• Standard Curve Preparation: Prepare the standard curve using serial dilutions of recombinant EGF in the same buffer or diluent as your samples to minimize matrix effects.

Limitations:

• If your recombinant EGF is not specifically labeled or validated for ELISA standard use, confirm its suitability by running parallelism and recovery experiments.
• Some recombinant proteins are formulated for bioassays rather than ELISA calibration; always check product documentation for intended use.

Summary Table: Recombinant Mouse EGF as ELISA Standard

In summary, recombinant mouse EGF is widely accepted and validated as a standard for ELISA quantification, provided it meets purity, activity, and validation criteria for your specific assay.

Recombinant Mouse EGF has been validated for multiple scientific applications in published research, primarily including bioassays, cell culture, functional assays, ELISA, Western blot, and immunohistochemistry.

Key validated applications and contexts:

• Bioassays: Used to assess mitogenic activity, cell proliferation, and differentiation in various cell types, including epithelial cells, fibroblasts, and stem cells.
• Cell Culture: Widely employed as a growth factor to support the expansion and maintenance of primary cells, stem cells (including neural and hematopoietic stem cells), organoids, spheroids, and tissue explants.
• Functional Assays: Validated for studies involving cell signaling, proliferation, migration, and differentiation, often through activation of downstream pathways such as RAS-RAF-MEK-ERK, PI3K-AKT, PLCγ-PKC, and STAT.
• ELISA and Western Blot: Used as a standard or control protein for quantification and detection of EGF or EGF-related signaling components.
• Immunohistochemistry: Applied for tissue localization and expression studies of EGF and its receptor.
• Tissue Dissociation: Utilized in protocols for dissociating tissues, particularly in neural and stem cell research.

Published research examples:

• Neurogenic reprogramming: Used in single-cell sequencing screens to identify compounds that enhance neurogenic reprogramming of murine Müller glia.
• Cancer biology: Applied in studies of ribosomal protein regulation in cancer cells, HER2+ metastatic breast cancer models, and prostate cancer progression.
• Stem cell biology: Supports the establishment and maintenance of pluripotent stem cells and epiblast-like stem cells.
• Immunology and metabolism: Investigated in models of immunosuppression in brain metastasis and metabolic switching in diabetic kidney cells.
• Organoid and spheroid culture: Facilitates growth and differentiation in organoid and spheroid models derived from mouse tissues.
• Wound healing and epithelial biology: Promotes proliferation and viability of epithelial cells and fibroblasts, relevant for tissue repair and regenerative medicine.

Additional notes:

• Recombinant Mouse EGF is a potent mitogen for many cell types and is frequently used in both in vitro and in vivo studies to induce proliferation and differentiation of skin, cornea, lung, trachea, and other tissues.
• It is also validated for use in tissue dissociation protocols, especially in neural and stem cell research.

These applications are supported by both product validation data and peer-reviewed publications, demonstrating the versatility of recombinant Mouse EGF in cell biology, stem cell research, cancer studies, and tissue engineering.

Reconstitution Protocol

Recombinant mouse EGF is supplied as lyophilized material and requires proper reconstitution before use in cell culture experiments. The standard reconstitution concentration is 0.1 mg/mL in sterile water, though some protocols recommend concentrations ranging from 0.1–1.0 mg/mL depending on your specific application.

Reconstitution Procedure

Pre-reconstitution steps:

• Centrifuge the vial at 3000 rpm for 5 minutes before opening to ensure all lyophilized material is at the bottom
• Allow the vial to reach room temperature if it has been stored at -20°C

Reconstitution technique:

• Add sterile water or sterile PBS to achieve your desired concentration
• Do not vortex the solution, as this can denature the protein
• Gently pipette and wash down the sides of the vial to ensure complete recovery of the protein into solution
• Allow several minutes for complete reconstitution

Storage and Stability

Short-term storage:Upon reconstitution, store the protein at 4°C for 2–7 days if you plan to use it within this timeframe.

Long-term storage:

• Store reconstituted aliquots at -80°C or below -18°C for extended periods
• For prolonged storage, dilute working aliquots in a 0.1% BSA (bovine serum albumin) or HSA (human serum albumin) solution to protect the protein from degradation
• Use a manual defrost freezer and avoid repeated freeze-thaw cycles, as these can compromise protein integrity

Further Dilution

After reconstitution, the protein can be further diluted into other aqueous solutions as needed for your specific cell culture applications. The choice of diluent should be based on your experimental requirements and assay conditions.