Recombinant Rat VEGF 165

Recombinant Rat VEGF 165 is widely used in research applications because it is a key regulator of angiogenesis, endothelial cell proliferation, and tissue repair, making it essential for studies involving vascular biology, tissue regeneration, and disease modeling.

Key scientific reasons to use Recombinant Rat VEGF 165:

• Angiogenesis and Endothelial Cell Proliferation: VEGF 165 is the most biologically active isoform of VEGF-A, stimulating the growth of new blood vessels and promoting proliferation of endothelial cells, which is critical for in vitro assays (e.g., HUVEC proliferation) and in vivo models of vascularization.

• Tissue Repair and Cytoprotection: Recombinant VEGF 165 has been shown to protect glomerular endothelial cells from injury and accelerate tissue repair, making it valuable for studies of renal disease, wound healing, and tissue regeneration.

• Disease Modeling: It is used to model pathological angiogenesis in cancer, cardiovascular disease, and ischemic conditions, allowing researchers to investigate mechanisms and potential therapeutic interventions.

• Species-Specific Activity: Using the rat isoform ensures optimal activity and compatibility in rat models, reducing cross-species variability and improving the relevance of experimental outcomes.

• Defined Biological Activity: Recombinant rat VEGF 165 is produced with high purity and specific activity, enabling reproducible and quantitative studies in cell-based assays and animal models.

Typical research applications include:

• Inducing angiogenesis in tissue engineering and regenerative medicine.
• Studying vascular responses in models of ischemia, wound healing, and organ repair.
• Investigating VEGF signaling pathways in normal physiology and disease states.
• Testing anti-angiogenic therapies in cancer and other diseases.

Best practices:
Use recombinant rat VEGF 165 at concentrations validated for your specific assay (e.g., HUVEC proliferation EC50 typically <8 ng/mL). Confirm species compatibility for in vivo studies to ensure physiological relevance. Validate biological activity in your system before experimental use.

In summary, Recombinant Rat VEGF 165 is a critical tool for vascular biology and regenerative research due to its potent, species-specific angiogenic and cytoprotective properties.

Yes, Recombinant Rat VEGF-165 can be used as a standard for quantification or calibration in ELISA assays designed to measure rat VEGF-165, provided that the ELISA kit or protocol is compatible with rat VEGF-165 and the recombinant protein is of high purity and well-characterized.

Key Points to Consider:

• Specificity: Recombinant Rat VEGF-165 is suitable as a standard for assays targeting rat VEGF-165. Most commercial rat VEGF ELISA kits (such as the Quantikine Rat VEGF ELISA Kit, RRV00) are calibrated using recombinant rat VEGF-165 or similar isoforms.
• Purity and Characterization: Ensure that the recombinant protein is highly purified and its concentration is accurately determined (e.g., by amino acid analysis or quantitative Western blot). This is critical for generating reliable standard curves.
• Compatibility: Confirm that the recombinant VEGF-165 is compatible with your ELISA system (e.g., capture and detection antibodies recognize the recombinant form).
• Formulation: Recombinant Rat VEGF-165 is typically supplied lyophilized and should be reconstituted according to the manufacturer’s instructions. It is often formulated in a buffer that does not interfere with ELISA assays.
• Stability: Store the recombinant protein at -20°C or lower, and avoid repeated freeze-thaw cycles to maintain activity.

Example from Literature and Kits:

• The Quantikine Rat VEGF ELISA Kit (RRV00) uses recombinant rat VEGF-165 as a standard.
• Many ELISA development kits (e.g., Invitrogen 900-K436) include recombinant rat VEGF-165 as the protein standard.

Recommendation:

If you are using a commercial ELISA kit, follow the manufacturer’s guidelines for standard preparation. If you are developing your own ELISA, use recombinant Rat VEGF-165 to generate a standard curve, ensuring that the protein is properly diluted in the same matrix as your samples (e.g., assay buffer or sample diluent).

In summary, Recombinant Rat VEGF-165 is appropriate and commonly used as a standard for quantifying rat VEGF-165 in ELISA assays, as long as it is of high quality and compatible with your assay system.

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

Validated Applications in Published Research:

• Functional Assays: Used to assess biological activity, such as stimulation of endothelial cell proliferation, migration, and tube formation, which are hallmarks of angiogenesis.
• ELISA (Enzyme-Linked Immunosorbent Assay): Applied as a standard or analyte to quantify VEGF levels in biological samples.
• Western Blot: Used as a positive control or to detect VEGF-related signaling pathways.
• Immunohistochemistry: Employed to visualize VEGF localization and expression in tissue sections.
• Immunoprecipitation: Utilized to study protein-protein interactions involving VEGF.
• Neurobiology Research: Investigated for its effects on the morphology and survival of neurons and glial cells, indicating roles beyond vascular biology.
• In Vivo Angiogenesis Models: Used in animal models to study vascular growth, tissue regeneration, and recovery from ischemic injury.
• Stem Cell Differentiation: Incorporated into protocols to promote differentiation of stem cells into endothelial or hematopoietic lineages, supporting vascular development in vitro.
• Retinal Vascular Studies: Applied in models of retinal ischemia to accelerate vascular recovery and reduce vaso-obliteration.
• Microparticle Delivery Systems: Used in conjugated or encapsulated forms to enhance cell survival and therapeutic efficacy in ischemic disease models.

Mechanistic and Activity Validation:

• Endothelial Cell Proliferation and Angiogenesis: Recombinant rat VEGF 165 acts via VEGFR-1 and VEGFR-2 receptors to modulate endothelial cell proliferation and angiogenesis, confirming its functional equivalence to endogenous VEGF.
• Neuronal and Glial Cell Survival: Research has validated its neurotrophic effects, expanding its utility to neurobiology.

Summary Table of Applications

These applications are supported by both direct validation in published research and by the established biological roles of VEGF 165 in angiogenesis, vascular biology, and neurobiology.

To reconstitute and prepare Recombinant Rat VEGF 165 protein for cell culture experiments, first centrifuge the vial briefly to collect the lyophilized powder at the bottom. Reconstitute the protein using sterile water at a concentration of 0.1 mg/mL (100 μg/mL). Gently pipette and wash down the sides of the vial to ensure complete dissolution and recovery of the protein.

Detailed protocol:

• Centrifuge the vial before opening to ensure all powder is at the bottom.
• Add sterile water to achieve a final concentration of 0.1 mg/mL (100 μg/mL).
• Gently pipette up and down and wash the vial walls to fully dissolve the protein.
• If long-term storage is required, add a carrier protein such as 0.1% human serum albumin (HSA) or bovine serum albumin (BSA) to the solution to enhance stability.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at –20°C or –80°C for long-term use.

Preparation for cell culture:

• Dilute the reconstituted stock solution into your cell culture medium to the desired working concentration, typically in the range of 4–8 ng/mL for endothelial cell stimulation.
• Ensure all solutions and diluents are sterile and endotoxin-free to prevent adverse effects on cell cultures.
• Avoid repeated freeze-thaw cycles by preparing single-use aliquots.

Additional notes:

• If your protocol or supplier recommends a specific buffer (e.g., phosphate-buffered saline, Tris-based buffer), use that instead of water.
• For optimal stability and activity, always consult the product-specific datasheet for any unique instructions regarding buffer composition or reconstitution conditions.

This approach ensures maximal recovery, stability, and biological activity of recombinant rat VEGF 165 for cell culture applications.