Recombinant Mouse VEGF 165

Recombinant Mouse VEGF 165 is widely used in research because it is a potent, well-characterized isoform of vascular endothelial growth factor (VEGF) that specifically stimulates angiogenesis, endothelial cell proliferation, and vascular permeability in mouse models. This makes it essential for studies involving vascular biology, tissue regeneration, cancer, and developmental biology.

Key reasons to use recombinant mouse VEGF 165 in research applications:

• Species specificity: Using the mouse isoform ensures optimal activity and receptor binding in mouse cells and tissues, minimizing cross-species variability and maximizing physiological relevance in murine models.
• Angiogenesis studies: VEGF 165 is the principal isoform responsible for promoting new blood vessel formation (angiogenesis), making it critical for investigating vascular development, wound healing, and tumor growth in vivo and in vitro.
• Endothelial cell activation: It robustly stimulates endothelial cell proliferation, migration, and survival, which are central processes in vascular biology and tissue engineering.
• Vascular permeability: VEGF 165 increases capillary permeability, allowing researchers to model and study vascular leakage, edema, and related pathologies.
• Signal transduction: It activates key signaling pathways via VEGF receptors (VEGFR1/Flt-1 and VEGFR2/Flk-1/KDR), enabling mechanistic studies of VEGF-mediated cellular responses.
• Regenerative medicine: VEGF 165 is used to enhance vascularization in engineered tissues and to study bone and organ regeneration due to its role in promoting blood supply to developing tissues.
• Disease modeling: It is essential for modeling diseases characterized by abnormal angiogenesis, such as cancer, diabetic retinopathy, and cardiovascular diseases.

Best practices:

• Use recombinant mouse VEGF 165 when working with mouse cells, tissues, or in vivo models to ensure biological relevance and reproducibility.
• Select the appropriate isoform (e.g., VEGF 165 vs. VEGF 121 or VEGF 189) based on the specific biological question, as different isoforms can have distinct effects on angiogenesis and tissue specificity.

In summary, recombinant mouse VEGF 165 is a critical reagent for any research focused on vascular biology, angiogenesis, and related disease or regenerative processes in mouse systems due to its potent, species-specific biological activity.

Yes, recombinant Mouse VEGF 165 can be used as a standard for quantification or calibration in ELISA assays, provided that the ELISA kit is designed to detect mouse VEGF and is compatible with recombinant standards.

Several ELISA kits for mouse VEGF (including VEGF 165) are calibrated using recombinant mouse VEGF standards, often expressed in systems such as Sf21 or E. coli. For example:

• The Mouse VEGF Quantikine ELISA Kit (R&D Systems, MMV00) is calibrated against a highly purified Sf21-expressed recombinant mouse VEGF164 (the murine equivalent of VEGF165), and it accurately quantitates both recombinant and natural mouse VEGF.
• Other commercial kits (such as those from PeproTech, Cell Sciences, and Arigo) also list recombinant mouse VEGF 165 as suitable for use as a standard.

Important considerations:

• Ensure the ELISA kit you are using is validated for mouse VEGF and specifies compatibility with recombinant standards.
• Confirm that the recombinant VEGF 165 you are using matches the isoform and sequence recognized by the kit’s antibodies.
• Follow the manufacturer’s instructions for preparing the standard curve, typically involving serial dilutions of the recombinant protein.

In summary, recombinant Mouse VEGF 165 is widely accepted and used as a standard for ELISA calibration and quantification of mouse VEGF in research settings.

Recombinant Mouse VEGF 165 has been validated for several key applications in published research, primarily related to its role in angiogenesis, endothelial cell biology, and functional assays.

Validated Applications:

• Functional Assays: Used to assess biological activity such as stimulation of endothelial cell proliferation, migration, and survival. It is commonly employed in in vitro assays to measure angiogenic potential and cell signaling responses.
• ELISA (Enzyme-Linked Immunosorbent Assay): Utilized as a standard or analyte to quantify VEGF levels in biological samples.
• Western Blot: Applied as a positive control or for detection of VEGF in protein extracts.
• Immunohistochemistry: Used to localize VEGF expression in tissue sections, often in studies of vascular development or pathology.
• Immunoprecipitation: Employed to isolate VEGF or its interacting partners from cell lysates.
• Cell Culture and Differentiation Studies: Supports the proliferation and migration of endothelial cells, and is used to induce angiogenic responses in various cell types.
• Chemoattractant Assays: Demonstrated to induce migration of monocytes and osteoblasts.
• Release of Endothelial Factors: Shown to increase the release of von Willebrand factor and metalloproteinase activity from endothelial cells.
• In Vivo Angiogenesis Models: Used in animal models to study neovascularization, vascular permeability, and tissue regeneration.

Supporting Details:

• VEGF 165 binds to VEGFR1 (FLT1) and VEGFR2 (KDR/Flk-1) receptors, activating downstream signaling pathways essential for angiogenesis and vascular permeability.
• It is frequently used in studies investigating hypoxia response, endothelial cell protection, and vascular development.
• Immobilized or microparticle-conjugated VEGF 165 has been used to enhance cell survival and therapeutic efficacy in ischemic disease models.
• Recombinant mouse VEGF 165 is suitable for SDS-PAGE analysis, confirming its purity and molecular weight for biochemical studies.

Summary Table of Applications

These applications are supported by both product validation data and peer-reviewed research, confirming the utility of recombinant mouse VEGF 165 in diverse experimental contexts.

To reconstitute and prepare Recombinant Mouse VEGF 165 protein for cell culture experiments, follow these steps for optimal solubility, stability, and biological activity:

1. Preparation Before Reconstitution

• Warm the lyophilized vial to room temperature before opening to minimize condensation and ensure complete recovery of the protein.
• Centrifuge the vial briefly (20–30 seconds) in a microcentrifuge to collect any protein on the cap or vial walls.

2. Reconstitution Buffer Selection

• For most cell culture applications, sterile water or low-pH buffer (e.g., 4 mM HCl) is recommended for initial reconstitution.
• If long-term stability or repeated freeze-thaw is a concern, add a carrier protein such as 0.1% human or bovine serum albumin (HSA/BSA) to the buffer.
• Avoid using buffers with high ionic strength or detergents unless specified for your application.

3. Reconstitution Procedure

• Add sterile water or buffer to achieve a final concentration of 100–500 μg/mL (not exceeding 1 mg/mL to avoid solubility issues).
• Gently swirl or tap the vial to dissolve the protein. Do not vortex or mix vigorously, as this may denature the protein.
• If solubility is incomplete, allow the solution to incubate at 4°C overnight to help dissolve aggregates.

4. Dilution for Cell Culture

• After reconstitution, further dilute the VEGF 165 stock solution into your cell culture medium to the desired working concentration (typically in the range of 1–100 ng/mL, depending on your assay requirements).
• If using for sensitive cell types or long-term storage, include 0.1% HSA/BSA in all dilutions to stabilize the protein.

5. Storage and Handling

• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can cause denaturation and loss of activity.
• Store aliquots at –20°C for long-term or 4°C for short-term (up to one week).
• Avoid storing at high concentrations (>1 mg/mL) and do not freeze/thaw repeatedly.

6. Quality Control

• Confirm protein recovery and integrity by running a small aliquot on SDS-PAGE; a band at the expected molecular weight (~39 kDa dimer) should be visible with as little as 10 ng loaded.

Summary Table: Mouse VEGF 165 Reconstitution

Additional Notes:

• Always use sterile, endotoxin-free reagents for cell culture applications.
• Adjust final working concentration based on your experimental design and cell type sensitivity.

These protocols ensure optimal solubility, stability, and biological activity of recombinant Mouse VEGF 165 for cell culture experiments.