Recombinant Mouse MSP R

Overview of Recombinant Mouse MSP R

Recombinant Mouse MSP R (macrophage stimulating protein receptor) is a valuable tool for research applications involving receptor signaling, cell migration, and immune function studies. This recombinant protein provides a well-characterized, reproducible system for investigating MSP-RON pathway biology in controlled experimental settings.

Key Research Applications

Binding and Activation Studies

Recombinant Mouse MSP R enables direct investigation of ligand-receptor interactions and receptor activation mechanisms. The protein can be used to study how macrophage stimulating protein (MSP) binds to and activates the receptor, facilitating dose-response and kinetic analyses. This is particularly useful for characterizing receptor dimerization and phosphorylation events that drive biological activity.

Cell Migration and Chemotaxis Assays

The MSP-RON pathway plays a critical role in cell migration and chemotaxis. Recombinant Mouse MSP R can be employed in functional assays to measure cellular responses to MSP stimulation, including macrophage migration and shape changes. These assays are essential for evaluating how receptor signaling influences cell motility and behavior in response to ligand engagement.

Neutralization and Blocking Studies

Recombinant Mouse MSP R serves as an effective target for antibody neutralization experiments, allowing researchers to assess the functional consequences of receptor inhibition. By combining the recombinant receptor with blocking antibodies, you can determine the specificity of antibody-receptor interactions and measure the degree of pathway inhibition in functional assays.

Biological Significance

The MSP-RON pathway regulates important physiological processes including inflammation modulation and tissue microenvironment stability. Recombinant Mouse MSP R provides a platform to study these regulatory mechanisms in vitro, offering insights into how receptor activation influences immune responses, metabolic processes, and tissue homeostasis without the complexity of whole-organism systems.

Practical Advantages

Recombinant proteins offer reproducibility and consistency across experiments compared to native protein sources. The availability of carrier-free formulations allows flexibility in experimental design, particularly when carrier proteins like bovine serum albumin would interfere with your specific application. Additionally, recombinant systems eliminate concerns about animal-derived pathogen contamination and provide standardized, well-characterized reagents suitable for quantitative research applications.

Yes, you can use Recombinant Mouse MSP R (Ron) Fc Chimera Protein as a standard for quantification or calibration in ELISA assays, but with some important considerations:

Key Points:

1. Appropriate Use as Standard:

   • Recombinant proteins, especially those specifically designed for immunoassays, are commonly used as standards to generate a standard curve for ELISA quantification. This allows you to convert your sample's optical density (OD) readings into absolute concentrations.

2. Critical Consideration - Analyte Match:

   • Ensure the ELISA kit is designed to detect Mouse MSP R (Ron). The standard must be the same molecule that the ELISA kit is measuring. If your ELISA kit is designed to detect Mouse MSP (MST1), then using Mouse MSP R (Ron) as a standard will not work, as they are different proteins (MSP is the ligand, MSP R/Ron is the receptor).
   • If your ELISA kit is specifically for Mouse MSP R (Ron), then the recombinant protein is suitable.

3. Formulation:

   • For use as an ELISA standard, it is generally recommended to use the recombinant protein formulated with BSA (Bovine Serum Albumin) rather than the carrier-free (CF) version. BSA helps stabilize the protein and reduces non-specific binding to the plate, leading to more accurate and reproducible standard curves.

4. Reconstitution and Dilution:

   • Follow the manufacturer's instructions for reconstituting the lyophilized protein.
   • Prepare a series of dilutions (e.g., 2-fold or 10-fold serial dilutions) in the same buffer used for your samples to create your standard curve. The range of your standard curve should bracket the expected concentration of your samples.

Summary:

Yes, you can use Recombinant Mouse MSP R Fc Chimera Protein as a standard for ELISA quantification, provided that:

• Your ELISA kit is designed to detect Mouse MSP R (Ron).
• You use the BSA-containing formulation for best results.
• You prepare the standard curve correctly according to the kit and protein manufacturer's guidelines.

If your ELISA kit detects Mouse MSP (MST1), you will need a Recombinant Mouse MSP (MST1) Protein as your standard instead.

Recombinant Mouse MSP R (Macrophage Stimulating Protein Receptor, also known as Ron or Stk) has been validated in published research for several key applications, primarily in studies of receptor-ligand interactions, cell signaling, and functional assays involving receptor activation and downstream biological effects.

Validated Applications in Published Research:

• Receptor Binding and Activation Assays:
  Recombinant Mouse MSP R has been used to study the binding of its ligand, MSP (Macrophage Stimulating Protein), and to assess receptor activation, including dimerization and phosphorylation events that trigger downstream signaling pathways. These assays are fundamental for understanding the biological activity of MSP and its receptor in various cell types.

• Functional Cell-Based Assays:
  The protein has been applied in cell-based experiments to evaluate biological responses such as cell migration, proliferation, and survival, particularly in epithelial cells and cells of the mononuclear phagocyte lineage. These assays help elucidate the physiological roles of MSP R in tissue repair, inflammation, and cancer biology.

• In Vitro Oligodendrocyte Studies:
  Recombinant MSP (and by extension, its receptor) has been used in vitro to investigate effects on oligodendrocyte process outgrowth and survival, especially in the context of demyelinating diseases. Application of recombinant MSP led to a reduction in oligodendrocyte process number, demonstrating its utility in neurobiology research.

• ELISA and Protein Quantification:
  Recombinant MSP R and its ligands are commonly used as standards or controls in ELISA assays to quantify protein concentrations and validate antibody specificity. Carrier-free forms are recommended when BSA could interfere with detection.

• Cancer and Biomarker Research:
  The receptor is a target in studies of cancer biomarkers, epithelial cell signaling, and receptor tyrosine kinase pathways, particularly those involving the Akt pathway. These applications often involve recombinant MSP R in both in vitro and in vivo models to dissect signaling mechanisms and therapeutic targeting.

Additional Context:

• Expression and Localization Studies:
  Recombinant MSP R has been used to probe its expression in various tissues, including the central and peripheral nervous systems, digestive tract, skin, lung, and immune cell populations.

• Neutralization and Blocking Studies:
  While not always using the recombinant receptor directly, antibodies against MSP R are validated for neutralization assays, which often employ recombinant protein as a target or competitor.

Summary Table of Validated Applications

If you need protocols or more specific details for any of these applications, please specify the experimental context.

To reconstitute and prepare Recombinant Mouse MSP (Macrophage Stimulating Protein, also known as MST1) R protein for cell culture experiments, follow these best-practice steps based on protocols for similar recombinant proteins and specific guidance for MSP:

1. Reconstitution

• Buffer: Use sterile phosphate-buffered saline (PBS) containing at least 0.1% carrier protein (such as bovine serum albumin, BSA, or human serum albumin) to minimize protein adsorption and loss.
• Concentration: A common working concentration for reconstitution is 100 μg/mL.
• Procedure:
  • Allow the lyophilized protein vial to equilibrate to room temperature before opening to prevent condensation.
  • Add the calculated volume of PBS + 0.1% BSA directly to the vial to achieve the desired concentration.
  • Gently swirl or invert the vial to mix. Avoid vigorous shaking or vortexing, as this can denature the protein.
  • Let the solution sit at room temperature for 15–30 minutes with gentle agitation to ensure complete dissolution.

2. Preparation for Cell Culture

• Sterility: Ensure all solutions and equipment are sterile to prevent contamination.
• Dilution: After reconstitution, further dilute the protein to the desired working concentration using cell culture medium (with serum or other acceptable carrier protein) immediately before use.
• Aliquoting: If not using the entire reconstituted solution at once, aliquot into single-use volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Storage: Store unused aliquots at –20 °C to –70 °C for long-term storage, or at 2–8 °C for up to one month after reconstitution under sterile conditions.

3. Additional Notes

• Carrier Protein: The presence of a carrier protein (e.g., BSA at 0.1–1%) is critical for stability, especially at low concentrations.
• Avoid Foaming: Do not vortex or pipette vigorously, as foaming can denature the protein.
• Documentation: Always consult the specific product datasheet for any unique requirements or recommendations.

Summary Table: Key Steps for Recombinant Mouse MSP R Protein Reconstitution

These steps will help ensure protein stability and bioactivity for your cell culture experiments. If your experiment requires a different buffer or concentration, adjust accordingly, but always maintain sterility and include a carrier protein for best results.