Recombinant Mouse EphB2

Recombinant Mouse EphB2 is a valuable tool for research applications involving cell signaling, neurobiology, cancer, fibrosis, and tissue regeneration due to its role as a receptor tyrosine kinase in the Eph/ephrin signaling pathway.

Key scientific applications and rationale:

• Neurobiology and CNS Regeneration: EphB2 regulates neuronal connectivity, migration of neural progenitor cells, neurogenesis, and synaptic plasticity. Exogenous recombinant EphB2 can be used to study these processes in vitro and in vivo, including models of spinal cord injury and cerebral infarction.

• Cancer Research: EphB2 is implicated in tumor angiogenesis, cell migration, and compartmentalization. Recombinant EphB2 can be used to investigate its role in tumor progression, metastasis, and as a potential therapeutic target.

• Fibrosis and Tissue Remodeling: EphB2 mediates dermal fibrosis and is involved in tissue compartmentalization. Recombinant protein enables mechanistic studies and therapeutic screening in models of systemic sclerosis and other fibrotic diseases.

• Immunology and Inflammation: EphB2 signaling influences neurogenic inflammation and enteric nervous system function, making recombinant EphB2 useful for assays in chronic colitis and other inflammatory conditions.

• Assay Development: Recombinant EphB2 is suitable for use in:

  • Cell-based assays (e.g., migration, proliferation, differentiation)
  • Immunoprecipitation and bioassays to study protein-protein interactions
  • In vivo models to assess physiological and pathological roles.

• Mechanistic Studies: The recombinant protein, often supplied as an Fc chimera, allows for controlled activation or blockade of EphB2 signaling, facilitating studies on bidirectional signaling with ephrin-B ligands.

Best practices:

• Use carrier-free preparations for binding and signaling studies to avoid interference.
• Select appropriate formats (e.g., Fc chimera) for enhanced stability and detection in assays.

Summary of scientific utility:
Recombinant Mouse EphB2 enables precise manipulation and analysis of EphB2-mediated signaling, supporting research in neural development, regeneration, cancer biology, fibrosis, and immune responses. Its use is essential for dissecting molecular mechanisms, validating therapeutic targets, and developing new intervention strategies in these fields.

Yes, recombinant Mouse EphB2 (such as the Fc Chimera Protein) can be used as a standard for quantification or calibration in ELISA assays, provided it is purified and its concentration is accurately determined.

Key Points:

• Suitability as Standard: Recombinant proteins, especially those that are purified and well-characterized (like the Recombinant Mouse EphB2 Fc Chimera), are commonly used as standards in ELISA assays to generate a standard curve for quantifying target protein concentrations in samples [2, 10, 11].
• Formulation: For ELISA standard use, it is generally recommended to use the recombinant protein with carrier (e.g., BSA) to improve stability and reduce non-specific binding, unless carrier-free is specifically required for your assay [10, 11].
• Concentration Determination: The concentration of the recombinant standard should be measured accurately, typically by methods such as HPLC or spectrophotometry, to ensure reliable quantification .
• Validation: The standard curve generated with recombinant Mouse EphB2 should be validated for linearity, sensitivity, and reproducibility in your specific ELISA setup .

Practical Considerations:

• Follow the manufacturer’s reconstitution and storage instructions for the recombinant protein.
• Ensure that the recombinant standard is compatible with the antibodies and detection system used in your ELISA.
• If possible, use a standard that has been validated for your specific application (e.g., ELISA) and species (mouse).

In summary, recombinant Mouse EphB2 is suitable for use as a standard in ELISA assays for quantification, as long as it is properly purified and its concentration is accurately known.

Recombinant Mouse EphB2 has been validated for several key applications in published research, primarily in studies of cell signaling, neurobiology, fibrosis, and immunology.

Validated Applications:

• In vitro cell signaling assays: Recombinant Mouse EphB2 is widely used to study EphB2-mediated forward signaling, including activation by ligands such as ephrin-B1 and Reelin, and subsequent downstream effects like cytoskeletal reorganization, cell retraction, and clustering in neuronal and non-neuronal cells.

• Neurobiology and axonal survival: Exogenous application of recombinant EphB2 protein has been shown to attenuate axonal degeneration in models of glaucoma, demonstrating its utility in neuroprotection and axonal survival assays.

• Fibrosis models: Recombinant EphB2 has been used to investigate its role in dermal fibrosis, particularly in systemic sclerosis (SSc) models. Studies have validated its involvement in fibroblast activation, collagen production, and skin thickening, both in vitro and in vivo.

• Cell adhesion and migration: EphB2 has been implicated in regulating monocyte adhesion and transendothelial migration, relevant for immunological studies and inflammation models.

• Angiogenesis and tumor biology: EphB2 carried on extracellular vesicles can induce angiogenesis by interacting with B-type ephrins on endothelial cells, supporting its use in angiogenesis and tumor microenvironment research.

• ELISA and Western blot standards: Recombinant Mouse EphB2 is validated as a standard for ELISA and Western blot assays, enabling quantification and detection of EphB2 in biological samples.

• Comparative immunology and preclinical therapeutic evaluation: The protein is used as a reagent in mouse model studies for comparative immunology and preclinical testing of therapeutic interventions targeting EphB2 signaling.

Additional Notes:

• Recombinant EphB2 is also used to study ligand-receptor specificity, receptor clustering, and downstream phosphorylation events in various cell types, including neuronal and fibroblast cultures.
• Some studies suggest roles in platelet activation, blood coagulation, and tumor suppression, though these applications are less frequently validated in mouse-specific recombinant protein research.

Summary Table:

These applications are supported by peer-reviewed studies and are considered validated uses for recombinant Mouse EphB2 in published research.

To reconstitute and prepare Recombinant Mouse EphB2 protein for cell culture experiments, centrifuge the vial briefly to collect the lyophilized powder, then add sterile distilled water to achieve a final concentration of 0.1–0.5 mg/mL. Avoid vortexing or vigorous pipetting to preserve protein integrity.

Step-by-step protocol:

• Centrifuge the vial before opening to ensure all powder is at the bottom.
• Add sterile distilled water to the vial to reach your desired concentration (typically 0.1–0.5 mg/mL for cell culture applications).
• Gently mix by slow pipetting or gentle inversion. Do not vortex, as this can denature the protein.
• Allow the protein to dissolve for several minutes at room temperature. If necessary, place the vial on a gentle shaker.
• Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at −20°C to −70°C for long-term storage, or at 2°C to 8°C for short-term use (up to one month).

Additional considerations:

• If your experiment requires a specific buffer (e.g., PBS), you may reconstitute directly in sterile PBS instead of water, but always check the product datasheet for buffer compatibility.
• For cell culture, ensure the final protein solution is sterile and endotoxin levels are appropriate for your application.
• If using the protein for functional assays, confirm activity post-reconstitution using a suitable assay.

Summary Table:

Always consult the specific product datasheet for any unique instructions or buffer requirements.