Recombinant Mouse VEGF 120

Recombinant Mouse VEGF 120 is widely used in research applications to study angiogenesis, vascular biology, and tissue regeneration because it is a key isoform of VEGF-A that stimulates endothelial cell proliferation, migration, and survival, while also influencing vascular permeability and vasodilation.

Key scientific reasons to use Recombinant Mouse VEGF 120:

• Angiogenesis Studies: VEGF 120 is a potent angiogenic factor, making it essential for experiments investigating blood vessel formation in vitro and in vivo.
• Endothelial Cell Function: It regulates the proliferation, migration, and survival of endothelial cells, which are critical for vascular development and repair.
• Vascular Permeability and Vasodilation: VEGF 120 can increase vascular permeability and stimulate vasodilation via nitric oxide-dependent pathways, relevant for studies on vascular physiology and pathology.
• Developmental Biology: It is expressed during embryogenesis and endochondral bone development, making it useful for research on skeletogenesis and tissue morphogenesis.
• Cancer Research: VEGF-A isoforms, including VEGF 120, are highly expressed in solid tumors and are central to tumor angiogenesis, supporting studies on cancer progression and anti-angiogenic therapies.
• Neurobiology and Regeneration: VEGF-A isoforms have roles in neuroprotection, axonal regeneration, and modulation of inflammation, relevant for studies on nervous system injury and repair.

Technical advantages of using the recombinant protein:

• Defined Isoform: VEGF 120 is a naturally occurring, alternatively spliced isoform of VEGF-A, lacking the heparin-binding domain present in longer isoforms (e.g., VEGF 164/165), which affects its diffusion and bioavailability in tissues.
• Consistency and Purity: Recombinant production ensures batch-to-batch consistency and high purity, which is critical for reproducible experimental results.
• Species Specificity: Mouse VEGF 120 is ideal for studies in murine models, ensuring optimal receptor binding and biological activity in mouse cells and tissues.

Typical research applications include:

• In vitro assays of endothelial cell proliferation, migration, and tube formation.
• In vivo models of angiogenesis, wound healing, and tissue regeneration.
• Studies of vascular permeability and inflammation.
• Investigation of VEGF signaling pathways in development, cancer, and neurobiology.

Best practices:
Use recombinant mouse VEGF 120 at concentrations optimized for your specific assay, and include appropriate controls to distinguish VEGF-specific effects from background activity. Always confirm the biological activity of the protein in your system, as isoform-specific responses may vary depending on cell type and experimental conditions.

Recombinant Mouse VEGF 120 can be used as a standard for quantification or calibration in ELISA assays, provided the assay is validated to recognize this isoform and the recombinant protein is of high purity and appropriate formulation. The suitability depends on the specificity of your ELISA antibodies and the standard curve preparation protocol.

Key considerations and supporting details:

• Isoform Recognition: Most mouse VEGF ELISA kits are designed to detect both the 120 and 164 amino acid residue forms of VEGF, as these are the major splice variants in mice. The assay documentation should confirm that the antibodies recognize VEGF 120, ensuring accurate quantification when using this isoform as a standard.

• Standard Curve Preparation: Quantitative ELISA requires a standard curve generated from known concentrations of the analyte. Recombinant mouse VEGF 120, if properly quantified and formulated, can be serially diluted to create this curve. Technical accuracy in preparation is essential for reliable results.

• Protein Purity and Formulation: The recombinant mouse VEGF 120 protein is typically supplied at >97% purity and is carrier-free or with BSA, depending on the application. Carrier-free preparations are preferred for ELISA standards to avoid interference. Ensure the protein is fully biologically active and properly stored to maintain integrity.

• Validation and Calibration: Some commercial ELISA kits calibrate against recombinant mouse VEGF 164, but their antibodies also recognize VEGF 120, and dose-response curves for both isoforms are parallel, allowing relative quantification. If your assay is calibrated against VEGF 164, using VEGF 120 as a standard is generally acceptable for relative quantification, but absolute values may differ slightly due to isoform-specific antibody affinities.

• Best Practices:

  • Confirm your ELISA kit’s documentation specifies recognition of VEGF 120.
  • Use the recombinant protein in the same buffer and diluent as recommended for standards.
  • Prepare a fresh standard curve for each assay run.
  • Validate the parallelism and recovery of VEGF 120 in your sample matrix to ensure assay accuracy.

• Limitations: If your ELISA kit is strictly validated for VEGF 164 or uses antibodies with isoform-specific affinity, results may not be directly comparable. Always consult the kit’s technical datasheet and, if necessary, perform a parallelism test to confirm equivalence.

In summary, recombinant mouse VEGF 120 is suitable as an ELISA standard if your assay recognizes this isoform and you follow validated protocols for standard curve preparation and calibration.

Recombinant Mouse VEGF 120 has been validated in published research primarily for applications involving angiogenesis assays, tissue engineering, and studies of vascular biology in murine models.

Key validated applications include:

• In vivo angiogenesis and tissue engineering models: Recombinant mouse VEGF 120 has been used to stimulate angiogenesis (formation of new blood vessels) in murine tissue engineering chamber models. For example, it was applied alone or in combination with other growth factors (such as FGF-2 and PDGF-BB) to assess its effects on angiogenesis and adipogenesis in engineered tissues. The study demonstrated that VEGF 120, especially when combined with FGF-2, synergistically increased angiogenesis at early time points in vivo.

• Cell culture and in vitro assays: VEGF 120 is commonly used in cell culture systems to promote endothelial cell survival, proliferation, migration, and tube formation, which are standard assays for evaluating angiogenic potential. It is also used as a positive control or standard in ELISA assays for VEGF quantification.

• Developmental biology and skeletogenesis: VEGF 120 has been validated in studies investigating its role in endochondral bone development and skeletogenesis, where it acts as a key angiogenic factor during these processes.

• Vascular permeability and endothelial function: Research has validated VEGF 120 for studying vascular permeability, endothelial cell survival, and migration, reflecting its physiological roles in vascular biology.

• Pathological angiogenesis models: VEGF 120 is used in models of tumor angiogenesis, wound healing, and ischemic injury to study pathological and reparative blood vessel formation.

Summary Table: Validated Applications of Recombinant Mouse VEGF 120

Additional notes:

• Recombinant mouse VEGF 120 is typically used in research settings and is not intended for diagnostic or therapeutic use in humans or animals.
• It is often compared or combined with other VEGF isoforms (such as VEGF 164/165) to dissect isoform-specific functions in angiogenesis and tissue remodeling.

If you require protocols or more specific assay details for a particular application, please specify the context (e.g., in vitro vs. in vivo, cell type, or disease model).

To reconstitute and prepare Recombinant Mouse VEGF 120 protein for cell culture experiments, follow these general best practices based on manufacturer protocols and scientific standards:

1. Centrifuge the vial before opening:
This ensures that all lyophilized protein is collected at the bottom and not lost when opening.

2. Reconstitution buffer:

• Use sterile PBS (phosphate-buffered saline, pH 7.2–7.4) or deionized water as the solvent, depending on the specific formulation and downstream application.
• For enhanced protein stability and to prevent adsorption to plastic, add 0.1% BSA (bovine serum albumin) to the buffer if compatible with your experiment.

3. Reconstitution concentration:

• A common working concentration is 50 μg/mL in sterile PBS.
• Alternatively, reconstitute to a concentration between 0.1–1.0 mg/mL in deionized water or buffered solution with 0.1% BSA for full activity.
• Adjust the volume according to the amount of protein and your experimental needs.

4. Gentle mixing:

• After adding the buffer, gently pipette up and down or swirl to dissolve the protein completely. Avoid vigorous vortexing to prevent denaturation.
• Wash down the sides of the vial to ensure full recovery of the protein.

5. Allow time for dissolution:

• Let the vial sit at room temperature for several minutes to ensure complete reconstitution.

6. Aliquot and storage:

• If not using immediately, aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at -80°C for long-term storage or at -20°C for short-term use. Avoid repeated freeze-thaw cycles.

7. Working solution preparation:

• Before adding to cell culture, dilute the stock solution to the desired final concentration using cell culture medium or PBS with 0.1% BSA, as appropriate for your assay.

Summary Table: Reconstitution Steps

Important notes:

• Always consult the Certificate of Analysis (CoA) or product datasheet for any protein-specific instructions, as formulation and recommended buffers may vary by supplier and tag.
• If the protein contains additives (e.g., BSA, trehalose), ensure compatibility with your downstream application.

These steps will help maintain the bioactivity and stability of recombinant mouse VEGF 120 for reliable cell culture experiments.