Recombinant Mouse VEGF R1

Recombinant mouse VEGF R1 (also known as Flt-1) offers several compelling advantages for research applications, particularly in studies involving vascular biology, angiogenesis, and disease modeling.

Key Research Applications

Angiogenesis and Vascular Development Studies

Recombinant mouse VEGF R1 serves as a critical tool for investigating vascular growth mechanisms. VEGF R1 binds VEGF with higher affinity than VEGF R2, making it particularly valuable for studying ligand-receptor interactions and signaling specificity. The receptor plays essential roles in neovascularization, wound healing, and vessel maintenance, allowing researchers to examine how VEGF signaling regulates these processes in controlled experimental settings.

Disease Modeling and Therapeutic Development

This recombinant protein is particularly useful for modeling pathological conditions. Research has demonstrated that VEGF R1 inhibition can improve muscle function and promote angiogenesis in muscular dystrophy models, suggesting therapeutic potential for degenerative diseases. By using recombinant mouse VEGF R1, you can develop and test therapeutic strategies targeting VEGF signaling pathways without the complications of whole-organism studies.

Functional Assays and Binding Studies

Recombinant mouse VEGF R1 is well-suited for multiple assay formats including bioassays, binding assays, and functional assays. The protein can be produced with various tags (such as Fc fusion proteins) to facilitate detection and purification, making it compatible with diverse experimental protocols and detection methods.

Regulatory Mechanism Investigation

VEGF R1 functions as a negative regulator of embryonic angiogenesis by inhibiting excessive endothelial cell proliferation. This makes the recombinant protein valuable for studying how cells balance pro-angiogenic signals and prevent pathological vascular overgrowth, relevant for understanding both normal development and disease states like cancer.

Practical Advantages

The availability of recombinant mouse VEGF R1 in various formats—including Fc-chimera proteins and different tag configurations—provides flexibility for your specific experimental needs. This standardization ensures reproducibility across experiments and facilitates comparison with published research protocols.

You can use recombinant mouse VEGF R1 as a standard for quantification or calibration in your ELISA assays, provided that the ELISA is specifically designed to detect mouse VEGF R1 and the recombinant protein is of high purity and well-characterized.

Key considerations and supporting details:

• ELISA Calibration: Quantitative ELISA assays require a standard curve generated from a known concentration of the target analyte, typically using a recombinant protein standard that matches the analyte being measured. For mouse VEGF R1, a recombinant mouse VEGF R1 protein is appropriate as a standard if the assay is validated for this purpose.

• Assay Compatibility: The standard must be recognized by the capture and detection antibodies in your ELISA. Commercial mouse VEGF R1 ELISA kits are calibrated using highly purified recombinant mouse VEGF R1 (often the soluble extracellular domain), and their protocols specify using this recombinant protein for the standard curve. Using a recombinant protein with the same sequence and post-translational modifications as the kit standard ensures accurate quantification.

• Validation: If you are using a custom or in-house ELISA, you must validate that your recombinant mouse VEGF R1 produces a standard curve that is parallel to those generated with natural or kit-provided standards. Parallelism indicates that the recombinant standard behaves similarly to the native protein in the assay, which is essential for accurate quantification.

• Concentration Range: Prepare a dilution series of the recombinant mouse VEGF R1 covering the expected concentration range in your samples, as described in the ELISA kit protocol.

• Interference: Ensure that the recombinant VEGF R1 does not interfere with other assay components or cross-react with other proteins in your samples. Some ELISAs may show interference at high concentrations of related proteins, so check the kit documentation for any such limitations.

Summary Table: Use of Recombinant Mouse VEGF R1 as ELISA Standard

If your recombinant mouse VEGF R1 meets these criteria, it is suitable for use as a standard in ELISA quantification and calibration. If using a commercial kit, always refer to the kit’s instructions and validation data to ensure compatibility.

Recombinant Mouse VEGF R1 (VEGFR1/Flt-1) has been validated in published research for a range of applications, primarily as a functional tool in bioassays, in vivo studies, ELISA standards, and binding assays.

Key validated applications include:

• Bioassays / Functional Assays: Used to study VEGF signaling, angiogenesis, and receptor-ligand interactions in cell-based and biochemical assays.
• In Vivo Studies: Applied in mouse models to investigate the role of VEGFR1 in vascular development, angiogenesis, inflammation, and disease models such as muscular dystrophy, sepsis, and bone healing.
• ELISA Standard: Utilized as a quantitative standard for VEGF or VEGFR1 in ELISA assays to measure protein levels in biological samples.
• Binding Assays: Employed to characterize the binding of VEGF or other ligands (including small molecules like TCDD) to VEGFR1, often using recombinant protein-coated plates.
• Protein Interaction Studies: Used in molecular docking and in vitro binding experiments to validate computational predictions of ligand-receptor interactions.

Representative published research applications:

• Bioassay/Functional Assay: Analysis of VEGF-induced autophosphorylation and downstream signaling in cell culture.
• In Vivo: Studies on the effects of VEGFR1 modulation in mouse models of angiogenesis, muscular dystrophy, sepsis, and bone formation.
• ELISA: Used as a standard for quantifying VEGF or VEGFR1 in biological fluids.
• Binding Assay: Validation of ligand (e.g., TCDD) binding to VEGFR1 using recombinant protein in vitro.

Additional context:

• VEGFR1 is a key regulator of angiogenesis, endothelial function, and inflammation, making recombinant forms valuable for dissecting these pathways in both basic and translational research.
• The recombinant protein is often used in both carrier-free and carrier-containing forms, depending on the application (e.g., cell culture vs. ELISA).

Summary Table of Validated Applications

These applications are well-supported in the literature and are considered standard for recombinant mouse VEGF R1 proteins in vascular biology and related fields.

To reconstitute and prepare Recombinant Mouse VEGF R1 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) to a concentration of 100 μg/mL, unless otherwise specified by the product datasheet. Use sterile technique throughout to maintain protein integrity and prevent contamination.

Detailed protocol:

• Reconstitution:

  • Allow the vial to equilibrate to room temperature before opening to minimize condensation.
  • Add sterile PBS (pH 7.2–7.4) directly to the vial to achieve a final concentration of 100 μg/mL (or as recommended by the datasheet; some protocols suggest a range of 0.1–1.0 mg/mL).
  • Gently swirl or invert the vial to mix. Do not vortex, as this can denature the protein.
  • Let the solution sit for 5–10 minutes at room temperature to ensure complete dissolution.

• Aliquoting and Storage:

  • Once fully dissolved, aliquot the protein into small volumes to avoid repeated freeze-thaw cycles, which can degrade protein activity.
  • Store aliquots at –20 °C to –70 °C, desiccated if possible, for long-term stability.
  • Avoid storing at 4 °C for extended periods, as this may reduce protein stability.

• Preparation for Cell Culture:

  • Before use, thaw an aliquot on ice.
  • If required, dilute further in sterile PBS or cell culture medium immediately before adding to cells.
  • For sensitive applications, consider adding 0.1% carrier protein (such as BSA or HSA) to minimize adsorption to plastic and improve stability, especially at low working concentrations.

Additional notes:

• Always consult the specific product datasheet for any unique instructions, as formulations and recommended reconstitution buffers may vary between preparations.
• If the protein is to be used in functional assays or bioactivity studies, confirm the absence of endotoxin or use a low-endotoxin preparation suitable for cell culture.
• Do not vortex or subject the protein to harsh agitation, as this can cause aggregation or loss of activity.

This protocol ensures the recombinant VEGF R1 protein is prepared in a manner compatible with cell culture experiments and preserves its biological activity.