Recombinant Mouse VEGF 164

Recombinant Mouse VEGF 164 is widely used in research because it is the principal isoform of mouse vascular endothelial growth factor A (VEGF-A), playing a critical role in angiogenesis, vascular permeability, endothelial cell survival, and neurogenesis. Its use enables precise control and reproducibility in studies investigating vascular biology, disease models, and therapeutic interventions.

Key scientific applications and advantages include:

• Angiogenesis Studies: VEGF 164 is a potent mitogen for vascular endothelial cells, making it essential for in vitro and in vivo models of blood vessel formation, wound healing, and tissue regeneration.
• Vascular Permeability and Endothelial Function: It regulates endothelial cell growth, migration, and barrier function, which are central to studies of vascular leakage, inflammation, and tumor biology.
• Disease Modeling: Elevated VEGF levels are associated with pathological conditions such as tumor angiogenesis, autoimmune diseases, diabetic retinopathy, and macular degeneration. Recombinant VEGF 164 allows for controlled investigation of these processes.
• Neurogenesis and Neuroprotection: VEGF 164 has neurotrophic effects, promoting the growth and survival of neurons and astrocytes, and is used in models of neurodegeneration and brain injury.
• Bioassays and Cell Culture: It is commonly used to stimulate endothelial cells in bioassays, enabling quantification of angiogenic responses and screening of anti-angiogenic compounds.
• Therapeutic Angiogenesis Research: Recombinant VEGF 164 is used to evaluate strategies for therapeutic vascularization, including gene delivery and tissue engineering.

Best practices for using recombinant mouse VEGF 164 include:

• Employing defined concentrations to mimic physiological or pathological conditions.
• Using carrier-free preparations for sensitive cell culture and bioassays.
• Validating activity in your specific cell type or animal model.

In summary, recombinant mouse VEGF 164 is a critical tool for dissecting mechanisms of vascular growth, permeability, and neurogenesis, and for modeling diseases where VEGF signaling is implicated.

Recombinant Mouse VEGF 164 can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and properly validated for this application.

For ELISA quantification:

• Recombinant Mouse VEGF 164 is commonly used as a standard in mouse VEGF ELISA kits. These kits are calibrated against highly purified recombinant mouse VEGF 164, and the standard curves generated with recombinant VEGF 164 are parallel to those obtained with natural mouse VEGF, allowing accurate quantification.
• Carrier proteins (such as BSA) are often included in recombinant VEGF preparations intended for use as ELISA standards to improve stability and reproducibility. Carrier-free forms are also available but may require additional handling precautions to prevent loss due to adsorption.
• The protein should be reconstituted and diluted according to the ELISA kit instructions, typically in the provided standard diluent buffer, to ensure compatibility and accurate calibration.

Best practices:

• Ensure the recombinant VEGF 164 is validated for ELISA use. Some recombinant proteins are specifically tested and certified for use as ELISA standards, while others are intended for bioassays or cell culture and may not be suitable for quantification.
• Check the purity and bioactivity. High purity (>95%) and confirmed bioactivity are important for reliable calibration.
• Prepare a standard curve using serial dilutions of the recombinant VEGF 164 in the same buffer as your samples to minimize matrix effects and ensure accurate quantification.

Limitations:

• For research use only: Recombinant VEGF 164 standards and ELISA kits are not intended for diagnostic procedures.
• Species specificity: Use mouse-specific antibody pairs for quantification of mouse VEGF, as cross-reactivity with other species may affect results.

In summary, recombinant mouse VEGF 164 is suitable as a standard for ELISA quantification if it is validated for this purpose, properly prepared, and used according to the assay protocol.

Recombinant mouse VEGF 164 has been validated for a diverse range of applications in published research, reflecting its critical role in vascular biology and tissue regeneration.

Primary Research Applications

Bioassay and Cell-Based Studies

The most extensively documented application is bioassay, with over 49 publications utilizing recombinant mouse VEGF 164 to assess angiogenic functions. These studies typically employ the protein to induce endothelial cell responses, including tube and cord formation assays in primary human retinal endothelial cells and human umbilical vein endothelial cells (HUVECs). The protein demonstrates potent bioactivity, with effective concentrations in the range of 0.5–3 ng/mL for inducing HUVEC cell proliferation.

Binding and Receptor Interaction Studies

Recombinant mouse VEGF 164 has been validated for binding assays and immunoprecipitation applications. These studies investigate the protein's interactions with its cognate receptors, VEGFR1 (Flt-1) and VEGFR2 (KDR/Flk-1), as well as its engagement with co-receptors neuropilin-1 (NRP-1) and neuropilin-2 (NRP-2). The protein's ability to bind heparan sulfate and remain retained on cell surfaces and in the extracellular matrix has been characterized through these binding assays.

Cell Culture and Differentiation

The protein has been applied in cell culture systems to study endothelial cell proliferation, migration, and survival. Additionally, it has been utilized in hematopoietic stem and progenitor cell research, as well as mammary epithelial cell research applications.

Specialized Research Applications

Tissue Engineering and Regeneration

Recombinant mouse VEGF 164 has been incorporated into engineered hydrogel systems for injured brain regeneration studies, demonstrating its utility in three-dimensional tissue engineering models. It has also been employed in tumor angiogenesis models using three-dimensional poly(ethylene glycol)-based systems to study vascular cell influences on tumor cell behavior.

In Vivo Studies

The protein has been validated for in vivo applications, including studies examining endothelial progenitor cell function in tumor xenograft models and lymphangiogenesis in animal models.

ELISA Applications

Recombinant mouse VEGF 164 serves as a standard and control reagent in enzyme-linked immunosorbent assays, with both capture and standard applications documented.

Signaling Pathway Analysis

Published research has utilized this protein to investigate downstream signaling cascades, including PI3K/AKT, p38 MAPK, FAK, and Paxillin activation pathways following VEGFR engagement. These studies have elucidated the molecular mechanisms underlying endothelial cell responses to VEGF 164 stimulation.

To reconstitute and prepare Recombinant Mouse VEGF 164 protein for cell culture experiments, dissolve the lyophilized protein at a concentration of 50 μg/mL in sterile PBS containing at least 0.1% serum albumin (either bovine or human). This helps stabilize the protein and maintain its biological activity.

Step-by-step protocol:

• Briefly centrifuge the vial before opening to ensure all contents are at the bottom.
• Add sterile PBS containing 0.1%–1% BSA (bovine serum albumin) or HSA (human serum albumin) to achieve a final concentration of 50 μg/mL. For some applications, higher concentrations (up to 0.1–1.0 mg/mL) may be used for stock solutions.
• Gently mix by pipetting up and down or by gentle vortexing. Avoid vigorous agitation or foaming.
• Allow the solution to sit at room temperature for 10–15 minutes to ensure complete dissolution.
• Aliquot the reconstituted protein to minimize freeze/thaw cycles, which can reduce activity.
• Store aliquots at ≤ –20°C for long-term storage. For short-term use, aliquots can be kept at 2–8°C for up to one week.

Additional notes for cell culture:

• For working dilutions, further dilute the stock solution in cell culture medium or buffer containing low endotoxin FBS or tissue culture grade BSA to maintain protein stability.
• Avoid repeated freeze/thaw cycles to preserve activity.
• Always use sterile technique to prevent contamination.

Summary of key points:

• Reconstitution buffer: Sterile PBS + 0.1%–1% BSA/HSA.
• Stock concentration: 50 μg/mL (or up to 1 mg/mL for some protocols).
• Aliquot and store: ≤ –20°C for long-term, 2–8°C for short-term.
• Avoid freeze/thaw cycles.
• Dilute in cell culture medium with carrier protein for experiments.

These steps ensure optimal solubility, stability, and biological activity of recombinant mouse VEGF 164 for cell culture applications.