Recombinant Mouse EphB4 is a valuable tool for research applications due to its well-characterized biological functions and its utility in studying EphB4 signaling pathways in mouse models. Here are several reasons why you should consider using Recombinant Mouse EphB4 in your research:

1. Study of EphB4 Signaling Pathways

Recombinant Mouse EphB4 allows for the direct investigation of EphB4-mediated signaling, including both forward (EphB4 receptor activation) and reverse (ephrinB2 ligand activation) signaling. This is critical for understanding cellular processes such as:

• Cell migration and adhesion
• Angiogenesis and vascular development
• Neurogenesis and neural repair
• Tumor growth and metastasis

2. Functional Assays

Recombinant Mouse EphB4 can be used in functional assays to:

• Assess the effects of EphB4 on cell proliferation, migration, and differentiation
• Investigate the impact of EphB4 on vascular integrity and remodeling
• Study the role of EphB4 in inflammatory responses and tissue repair

3. Therapeutic Target Validation

EphB4 is a potential therapeutic target in various diseases, including cancer and ischemic stroke. Using recombinant Mouse EphB4, you can:

• Validate the efficacy of EphB4-targeting therapies
• Study the mechanisms of action of EphB4 inhibitors or activators
• Evaluate the effects of EphB4 modulation on disease progression

4. Control and Standardization

Recombinant proteins provide a consistent and standardized source of EphB4, which is essential for:

• Reproducibility of experiments
• Comparison of results across different studies
• Quality control in high-throughput screening

5. In Vivo and In Vitro Studies

Recombinant Mouse EphB4 can be used in both in vitro and in vivo studies, allowing for:

• Cell culture experiments to study EphB4 signaling
• Animal models to investigate the physiological and pathological roles of EphB4

6. Cross-Species Compatibility

Recombinant Mouse EphB4 is compatible with mouse models, making it ideal for studies that require species-specific interactions and signaling.

7. Support for Antibody and Assay Development

Recombinant Mouse EphB4 can be used as a positive control in immunoassays and for the development and validation of antibodies and other detection reagents.

8. Research on Disease Mechanisms

EphB4 has been implicated in various diseases, including:

• Cancer (tumor growth, angiogenesis, and metastasis)
• Cardiovascular diseases (vascular integrity and remodeling)
• Neurological disorders (neurogenesis and neural repair)

Using recombinant Mouse EphB4, you can gain insights into the molecular mechanisms underlying these conditions and develop new therapeutic strategies.

Summary

Recombinant Mouse EphB4 is a versatile and essential tool for researchers studying EphB4 signaling, disease mechanisms, and therapeutic interventions. Its use ensures consistency, reproducibility, and the ability to perform both in vitro and in vivo experiments, making it a valuable asset in your research applications.

Yes, you can use Recombinant Mouse EphB4 as a standard for quantification or calibration in your ELISA assays, provided it is purified and its concentration can be accurately determined.

Key Points for Using Recombinant Mouse EphB4 as an ELISA Standard:

1. Purified Protein: Recombinant Mouse EphB4 (especially in carrier-free form) is typically purified and suitable for use as a standard. The protein should be free of contaminants that could interfere with ELISA detection.

2. Quantitative Accuracy: For accurate quantification, the concentration of the recombinant protein must be measured precisely (e.g., by amino acid analysis, HPLC, or spectrophotometry). Many commercial suppliers provide the protein with a certificate of analysis specifying concentration and purity.

3. Standard Curve Preparation: Prepare a serial dilution of the recombinant EphB4 to generate a standard curve. The range should cover the expected concentrations in your samples (commonly 0–1000 pg/mL or higher, depending on your assay sensitivity).

4. Carrier Protein Consideration: If the recombinant EphB4 is supplied with a carrier protein (like BSA), this may affect the assay if the carrier interferes with detection or binding. For ELISA calibration, carrier-free (CF) versions are generally preferred.

5. Reconstitution and Storage: Follow the manufacturer’s instructions for reconstitution and storage. Lyophilized proteins should be reconstituted in the recommended buffer and aliquoted to avoid repeated freeze-thaw cycles.

6. Validation: Validate the standard curve in each assay run to ensure linearity and reproducibility. The correlation coefficient (R²) of the standard curve should be ≥0.98 for reliable quantification.

References Supporting Use as ELISA Standard:

• Recombinant Mouse EphB4 has been used in ELISA assays for quantification and calibration (e.g., in studies cited in R&D Systems and Leinco product datasheets).
• General guidelines recommend using purified recombinant proteins as standards for ELISA (Boster Bio, Assay Genie, FineTest, GENLISA, XpressBio).

Summary:

Yes, Recombinant Mouse EphB4 is appropriate for use as a standard in ELISA assays for quantification or calibration, as long as it is purified, accurately quantified, and used according to best practices for standard curve generation.

Applications of Recombinant Mouse EphB4 in Published Research

Recombinant Mouse EphB4 has been validated across multiple research applications, with particular emphasis on functional and signaling studies. The primary applications documented in published research include:

Bioassay and Functional Studies

Bioassay represents the most extensively documented application for recombinant Mouse EphB4. This protein has been utilized to investigate EphB4/ephrinB2 signaling in various biological contexts, including vascular remodeling, angiogenesis, and bone metabolism. Specifically, recombinant EphB4-Fc chimera constructs have been employed to study activation of ephrinB2 signaling in murine arteriovenous fistulae and to examine the effects of EphB4 on osteoclast function and bone turnover.

Cellular and Molecular Analysis

The protein has been validated for use in whole cell assays and cell lysate analysis, enabling researchers to assess EphB4 signaling pathway activation and downstream effects on cell proliferation and differentiation. Research has demonstrated that recombinant EphB4 can modulate the ABL1/Cyclin D1 signaling pathway, promoting neural stem cell proliferation and neuronal differentiation following ischemic injury.

Immunological Applications

Recombinant Mouse EphB4 has been used as a capture reagent in ELISA assays and for immunoprecipitation studies, facilitating the detection and quantification of EphB4-binding partners and signaling intermediates. These applications support both protein-protein interaction studies and quantitative analysis of EphB4 expression levels.

In Vivo Research

The protein has been validated for in vivo applications, where recombinant EphB4-Fc constructs have been administered to investigate vascular repair mechanisms, neuroinflammation reduction, and therapeutic potential in disease models such as cerebral ischemia and myeloma-associated bone disease.

These validated applications demonstrate that recombinant Mouse EphB4 serves as a versatile tool for investigating Eph receptor signaling in both basic research and translational studies.

To reconstitute and prepare Recombinant Mouse EphB4 protein for cell culture experiments, dissolve the lyophilized protein at a concentration of 100 μg/mL in sterile PBS. This concentration is commonly used for bioassays and cell-based applications.

Step-by-step protocol:

• Before opening the vial: Briefly centrifuge to collect all lyophilized material at the bottom.
• Reconstitution: Add sterile PBS directly to the vial to achieve a final concentration of 100 μg/mL. Gently agitate and allow the protein to dissolve for 15–30 minutes at room temperature; avoid vigorous shaking to prevent foaming and protein denaturation.
• Sterility: Use sterile technique throughout to prevent contamination.
• Aliquoting: Once fully dissolved, aliquot the solution to minimize freeze-thaw cycles, which can degrade protein activity.
• Storage: Store aliquots at −20 °C or −80 °C in a manual defrost freezer. Avoid repeated freeze-thaw cycles for optimal stability.
• Working solution: For cell culture, dilute the stock solution further in cell culture medium as required for your experimental design. Ensure the final buffer composition is compatible with your cells.

Additional notes:

• If your protocol or datasheet specifies a different buffer (e.g., sterile water or another buffer), follow those instructions, but PBS is standard for EphB4.
• For sensitive cell types, confirm that the buffer and any carrier proteins (e.g., BSA) are compatible with your assay.
• Always consult the specific product datasheet for any unique instructions regarding your batch of recombinant EphB4.

This approach ensures the protein is properly reconstituted and maintains biological activity for cell culture experiments.