Recombinant Human EGF R

Recombinant Human Epidermal Growth Factor (rhEGF) is a powerful and versatile tool for a wide range of research applications due to its well-characterized biological activity, reproducibility, and purity. Here are several compelling reasons to use rhEGF in your research:

1. High Bioactivity and Consistency

Recombinant EGF is produced under controlled conditions, ensuring high bioactivity, batch-to-batch consistency, and purity. This is critical for reproducible experimental results, especially in sensitive assays and cell culture applications.

2. Promotes Cell Proliferation and Viability

rhEGF is a potent mitogen that stimulates the proliferation and viability of various cell types, including epithelial cells (e.g., HaCaT), fibroblasts (e.g., NIH 3T3), and stem cells. It activates key signaling pathways such as ERK1/2 and c-Jun, which regulate cell growth and survival.

3. Supports Wound Healing and Tissue Regeneration

rhEGF plays a central role in wound healing by promoting epithelial cell migration, proliferation, and tissue remodeling. It is widely used in studies of skin repair, chronic wound models, and tissue engineering.

4. Enables Organoid and 3D Cell Culture

Recombinant growth factors like EGF are essential for the development and maintenance of organoids and 3D cell cultures. These models more accurately mimic in vivo tissue architecture and function, making them valuable for drug screening, disease modeling, and regenerative medicine research.

5. Facilitates Stem Cell Research

rhEGF supports the proliferation and differentiation of stem cells, including those in the intestine and central nervous system. It is a key component in protocols for directed differentiation and expansion of stem cell populations.

6. Reduces Experimental Variability

Compared to native or animal-derived growth factors, recombinant EGF offers superior purity and reduced risk of contamination (e.g., endotoxins, viruses). This minimizes experimental variability and enhances the reliability of your results.

7. Broad Applicability Across Research Fields

rhEGF is used in diverse areas such as:

• Dermatology: Wound healing, skin aging, acne, and radiation dermatitis.
• Cancer Research: Studying tumor growth, angiogenesis, and cell signaling.
• Cardiovascular Research: Investigating blood vessel formation and tissue repair.
• Neurobiology: Supporting neuron survival and synaptic function.
• Environmental and Agricultural Research: Studying plant hormone signaling and biodegradation.

8. Well-Documented and Cited

rhEGF is extensively cited in the scientific literature and has been validated in numerous peer-reviewed studies, providing confidence in its performance and reliability.

9. Animal-Free and Endotoxin-Free Options Available

Many recombinant EGF products are produced in animal-free systems and are free of endotoxins, making them suitable for sensitive cell culture and clinical research applications.

10. Supports Mechanistic Studies

rhEGF allows researchers to dissect the molecular mechanisms underlying cell proliferation, differentiation, and tissue regeneration, including the regulation of gene expression, protein phosphorylation, and signaling pathway activation.

In summary, recombinant Human EGF is a reliable, potent, and versatile growth factor that enhances the quality and reproducibility of your research across multiple disciplines.

Recombinant Human EGF R (Epidermal Growth Factor Receptor) can be used as a standard for quantification or calibration in ELISA assays, provided it is well-characterized, pure, and compatible with the antibodies and assay format.

Key considerations and supporting details:

• ELISA kits for EGFR quantification routinely use recombinant human EGFR as the standard to generate calibration curves for quantifying EGFR in biological samples. The recombinant protein is serially diluted to create a standard curve, and sample concentrations are interpolated from this curve.
• Assay validation data show that recombinant EGFR spiked into various matrices (cell culture media, plasma, serum) yields high recovery rates (typically 98–101%), indicating suitability for use as a standard.
• Critical requirements for using recombinant EGFR as a standard:
  • The recombinant protein must be of high purity and accurately quantified.
  • The protein should match the epitope recognized by the capture and detection antibodies in your ELISA.
  • The standard curve should be linear across the assay’s dynamic range, and parallelism between standard and sample curves should be confirmed to ensure accurate quantification.
• Protocol best practices:
  • Reconstitute and dilute the recombinant EGFR according to the ELISA kit instructions, using the recommended calibrator diluent for your sample type.
  • Store the recombinant standard at -20 °C or -80 °C to maintain stability.
  • Validate the assay by assessing recovery, linearity, and parallelism with your specific sample matrix.

Limitations:

• The recombinant EGFR standard should be validated for your specific ELISA system, as differences in protein isoforms, glycosylation, or tags may affect antibody recognition and quantification accuracy.
• For regulatory or diagnostic applications, reference standards calibrated to international units (e.g., NIBSC) may be required.

Summary: If your recombinant human EGFR is well-characterized and matches the assay’s antibody specificity, it is appropriate for use as a standard in ELISA quantification and calibration. Always validate performance in your specific assay context.

Recombinant human epidermal growth factor (rhEGF) has been validated for numerous applications across multiple research domains, as demonstrated in published literature.

Cell Proliferation and Viability Studies

rhEGF is extensively used to promote cell proliferation and enhance cell viability across different cell types. Research has demonstrated that purified recombinant EGF effectively stimulates the proliferation of fibroblasts and epithelial cells, with significant increases in cell viability observed following treatment. The protein activates intracellular signaling pathways, particularly the ERK1/2 and c-Jun signaling cascades, which upregulate expression of proteins critical for cell proliferation such as PCNA, HMGA1, and MCM2.

Signaling Pathway Analysis

rhEGF serves as a tool for investigating EGF receptor signaling mechanisms. The protein activates at least four major downstream signaling cascades: the RAS-RAF-MEK-ERK pathway, PI3 kinase-AKT pathway, PLCgamma-PKC pathway, and STAT modules. This makes it valuable for studying receptor activation and downstream molecular events in various cellular contexts.

Tissue Engineering and Regenerative Medicine

rhEGF has been validated for applications in wound healing and tissue repair. Exogenous administration of bioidentical human recombinant epidermal growth factor has demonstrated efficacy in promoting skin wound healing and minimizing scar formation. Additionally, EGF stimulates cell proliferation and differentiation in the intestine and promotes proliferation of stem cells in the central nervous system.

Assay Development

rhEGF is validated for use in multiple assay formats including ELISA, blocking assays, functional assays, and Western blot applications. These applications support both basic research and therapeutic development efforts.

Gene Expression and Functional Studies

RNA sequencing analysis has revealed that rhEGF increases transcription of genes enriched in ribosome biogenesis and cell proliferation pathways. Gene set enrichment analysis demonstrates that rhEGF promotes multiple biological functions including DNA replication, metabolic processes, mRNA processing, and protein folding.

To reconstitute and prepare Recombinant Human EGF R protein for cell culture experiments, dissolve the lyophilized protein in sterile, endotoxin-free buffer—commonly sterile PBS (pH 7.4) or distilled water—at a recommended concentration specified in the product documentation, typically ranging from 10 μg/mL to 0.2 mg/mL.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Add sterile buffer: Use sterile PBS (pH 7.4) or distilled water. For enhanced stability and to prevent adsorption, include 0.1% human or bovine serum albumin (HSA/BSA) if recommended.
• Reconstitution concentration: Common working concentrations are 10 μg/mL, 0.2 mg/mL, or as specified in the Certificate of Analysis (CoA) for your batch. Adjust volume accordingly.
• Mix gently: Swirl or tap the vial to dissolve the protein. Avoid vigorous shaking to prevent foaming and protein denaturation.
• Allow to dissolve: Let the solution sit at room temperature for 15–30 minutes with gentle agitation.
• Aliquot: Divide into small working aliquots to avoid repeated freeze-thaw cycles, which can denature the protein.
• Storage: Store reconstituted protein at 4°C for short-term use (up to 1 month), or at –20°C to –80°C for long-term storage (up to 3 months). Avoid frost-free freezers and repeated freeze-thaw cycles.

Additional notes:

• Always consult the product’s Certificate of Analysis (CoA) or technical datasheet for batch-specific instructions, as optimal buffer and concentration may vary.
• If a precipitate forms, microcentrifuge briefly before use to clarify the solution.
• For cell culture, ensure all reagents are endotoxin-free to prevent adverse cellular responses.
• For functional assays, typical working concentrations range from 0.5–25 ng/mL, but titrate as needed for your specific cell type and application.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of recombinant EGF R protein for cell culture experiments. Always verify with your specific product documentation for any unique requirements.