Recombinant Human EG-VEGF

Recombinant Human EG-VEGF (Endocrine Gland-derived Vascular Endothelial Growth Factor, also known as prokineticin-1 or PK1) is a specialized angiogenic growth factor with distinct biological roles compared to classical VEGFs. You should consider using recombinant human EG-VEGF in your research applications if your work involves studying tissue-specific angiogenesis, particularly in endocrine organs, or if you are investigating mechanisms of vascularization, cell signaling, or tissue regeneration in these contexts.

Key reasons to use Recombinant Human EG-VEGF in research:

• Tissue-Specific Angiogenesis: EG-VEGF is highly expressed in the ovaries, testis, adrenal glands, and placental tissues, where it regulates blood vessel formation and function in a manner distinct from VEGF-A. This makes it valuable for studying angiogenesis in reproductive and endocrine tissues.

• Cellular and Molecular Mechanisms: EG-VEGF acts through specific receptors (PROKR1 and PROKR2), enabling research into unique signaling pathways that differ from those activated by classical VEGFs. This is important for dissecting the molecular basis of tissue-specific vascularization and for identifying novel therapeutic targets.

• Disease Modeling: Dysregulation of EG-VEGF has been implicated in reproductive disorders, tumor angiogenesis in endocrine tissues, and placental pathologies. Recombinant EG-VEGF can be used to model these diseases in vitro or in vivo, facilitating the study of pathophysiological mechanisms and the development of targeted therapies.

• Assay Development: Recombinant EG-VEGF is suitable for use as a standard in ELISA, for cell stimulation in culture, and for functional assays such as proliferation, migration, and tube formation of endothelial cells derived from endocrine tissues.

• Bioprocessing and Cell Therapy: EG-VEGF can be used in cell culture systems to optimize the growth and differentiation of cells from endocrine organs, or in bioprocessing applications where tissue-specific vascularization is required.

Best practices:

• Use carrier-free preparations for sensitive applications or when downstream purification is required.
• Select formulations with stabilizers (such as BSA) for routine cell culture or as assay standards to enhance protein stability.

In summary: Use recombinant human EG-VEGF when your research requires a precise, tissue-specific angiogenic factor relevant to endocrine organ biology, disease modeling, or specialized vascularization studies. Its unique receptor specificity and biological activity make it a powerful tool for dissecting angiogenic mechanisms beyond those addressed by classical VEGFs.

You can use recombinant human EG-VEGF as a standard for quantification or calibration in your ELISA assays, provided it is compatible with your assay system and matches the analyte detected by your ELISA kit. Most commercial EG-VEGF ELISA kits use recombinant human EG-VEGF as the standard for generating the calibration curve.

Key considerations:

• Standard Curve Preparation: ELISA kits for EG-VEGF typically require the preparation of a standard curve using serial dilutions of recombinant human EG-VEGF, with detection ranges commonly between 15.6–1000 pg/mL or similar. The standard should be reconstituted and diluted according to the kit instructions to ensure accuracy.

• Assay Compatibility: The recombinant EG-VEGF used as a standard should be of the same isoform and sequence as the target analyte in your samples. Most kits specify the use of recombinant human EG-VEGF (also known as PROK1) for calibration.

• Validation: The standard curve must be run with each assay to ensure reliable quantification. The kit protocol will specify the required concentrations and dilution steps.

• Matrix Effects: If you are quantifying EG-VEGF in complex biological samples (e.g., serum, plasma, tissue lysates), ensure that the recombinant standard behaves similarly to endogenous EG-VEGF in your sample matrix. Some kits recommend performing spike-and-recovery or parallelism experiments to confirm this.

• Documentation: Always refer to your specific ELISA kit’s manual for detailed instructions on standard preparation and calibration, as protocols may vary between manufacturers.

Summary Table: Use of Recombinant Human EG-VEGF as ELISA Standard

Conclusion:
Recombinant human EG-VEGF is widely accepted and routinely used as a standard for quantification and calibration in EG-VEGF ELISA assays, as long as it is compatible with your assay system and matches the analyte detected by your kit. Always follow your kit’s protocol for best results.

Recombinant Human EG-VEGF (Endocrine Gland-Derived Vascular Endothelial Growth Factor, also known as PK1 or PROK1) has been validated in published research primarily for applications involving angiogenesis assays, cell proliferation, migration studies, and as a functional standard in biochemical assays.

Key validated applications in published research include:

• Cell-based functional assays: EG-VEGF has been used to stimulate proliferation, migration, and tube formation in endothelial cells, particularly those derived from microvascular beds of endocrine tissues such as placenta, ovary, and testis. These assays are commonly used to assess angiogenic potential and signaling activity.

• Biochemical and analytical assays: Recombinant EG-VEGF is validated for use as a standard in ELISA (enzyme-linked immunosorbent assay) and HPLC (high-performance liquid chromatography), as well as for SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) to confirm purity and molecular weight.

• Cancer research: EG-VEGF has been studied for its role in tumor angiogenesis and its deregulation in cancers such as colorectal cancer, where it is used to investigate mechanisms of tumor vascularization and potential therapeutic targeting.

• Tissue and organ-specific angiogenesis studies: EG-VEGF is particularly relevant in research on the vascularization of endocrine organs (placenta, ovary, testis, adrenal gland), where it is used to model and dissect tissue-specific angiogenic pathways.

• Cell culture supplement: It is used as a growth factor in cell culture systems to support the growth and maintenance of specific endothelial cell types or to induce angiogenic responses in vitro.

Summary Table of Validated Applications

Notes:

• EG-VEGF is distinct from classical VEGF-A and is primarily active in microvascular endothelial cells of endocrine tissues.
• Most published research focuses on its role in angiogenesis, cancer biology, and as a tool in cell-based and biochemical assays.

If you require protocols or more detailed examples from specific studies, please specify the application area of interest.

To reconstitute and prepare Recombinant Human EG-VEGF (Endocrine Gland-Derived Vascular Endothelial Growth Factor) for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Open the vial carefully under sterile conditions.

Reconstitution:

• Add sterile water or the recommended buffer to achieve a final concentration of 0.1 mg/mL (100 μg/mL) unless your protocol or datasheet specifies otherwise.
• Gently pipette the solution up and down along the vial wall to dissolve the protein completely. Avoid vigorous mixing or vortexing, as this can denature the protein.
• Let the vial sit at room temperature for 10–30 minutes to ensure full dissolution.
• If necessary, gently swirl or invert the vial, but do not create foam.

Aliquoting and Storage:

• Once fully dissolved, aliquot the solution into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at –20°C to –70°C for long-term storage. For short-term use (up to 1 month), store at 2–8°C under sterile conditions.

Preparation for Cell Culture:

• Before adding to cell cultures, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or buffer compatible with your assay.
• If the protein was reconstituted in water, ensure the final buffer composition is compatible with your cells (e.g., isotonic, pH 7.2–7.4, free of toxic additives).

General Notes:

• Always consult the specific product datasheet for any unique requirements (e.g., use of carrier proteins like BSA, or specific buffer recommendations).
• Avoid repeated freeze-thaw cycles by aliquoting.
• Use sterile technique throughout to prevent contamination.

Summary Table: EG-VEGF Reconstitution Protocol

These steps ensure optimal recovery, stability, and biological activity of recombinant EG-VEGF for cell culture applications.