Recombinant Human MSP

Recombinant Human MSP (Macrophage Stimulating Protein) is used in research applications to study its roles in cell signaling, immune modulation, tissue regeneration, and disease mechanisms, offering high purity, consistency, and functional activity for experimental reproducibility.

Key scientific reasons to use Recombinant Human MSP:

• Cellular Function Studies: MSP is a growth factor that induces proliferation of macrophages and keratinocytes, activates osteoclasts, and exerts anti-apoptotic effects, making it valuable for investigating cell signaling pathways and tissue regeneration processes.

• Immunomodulation Research: MSP inhibits LPS- or IFN-induced iNOS and IL-12 expression, implicating it in the regulation of inflammatory responses and immune cell function. This is critical for studies on innate immunity, inflammation, and autoimmune disease models.

• Disease Mechanism Elucidation: MSP’s involvement in tubular regeneration after acute kidney injury and its proliferative/anti-apoptotic effects are relevant for research into kidney disease, wound healing, and tissue repair.

• Drug Discovery and Screening: Recombinant MSP can be used in high-throughput assays to identify compounds that modulate its activity, supporting drug target validation and therapeutic development.

• Structural and Biochemical Analysis: Recombinant MSP provides sufficient material for structural biology techniques (e.g., X-ray crystallography, NMR), enabling detailed studies of its molecular interactions and binding sites.

• Experimental Consistency: Recombinant proteins offer batch-to-batch reproducibility, defined sequence, and high purity, minimizing variability and ensuring reliable results in functional assays and mechanistic studies.

• In Vitro and In Vivo Applications: MSP can be used in cell culture systems to study its direct effects on target cells, or in animal models to investigate its physiological and pathological roles.

Best practices for using Recombinant Human MSP:

• Use tag-free preparations for in vivo studies to minimize immunogenicity and ensure native protein function.
• Validate biological activity in your specific assay system, as MSP’s effects may be context-dependent.
• Employ appropriate controls, such as untreated cells or cells treated with inactive protein, to distinguish MSP-specific effects.

Summary of scientific applications:

• Cell signaling and proliferation assays
• Immune modulation studies
• Tissue regeneration and repair models
• Drug screening and target validation
• Structural biology and protein interaction analysis

Recombinant Human MSP is a versatile tool for dissecting its biological roles and therapeutic potential, with advantages in purity, consistency, and experimental control that are essential for rigorous scientific research.

Yes, you can use Recombinant Human MSP as a standard for quantification or calibration in your ELISA assays, provided it is suitable for your specific ELISA format and antibody pair.

Key Points:

• Suitability for ELISA: Recombinant Human MSP proteins (such as those from Abcam, Novus Biologicals, or R&D Systems) are commonly used as standards in sandwich ELISA assays. For example, Abcam’s Recombinant Human MSP Protein Standard (His tag) is explicitly stated to be suitable for sELISA and is guaranteed to work with their capture and detector antibodies.

• Calibration and Quantification: To use recombinant MSP as a standard, you must generate a standard curve by serially diluting the recombinant protein and measuring its signal in your ELISA. This curve allows you to interpolate the concentration of MSP in your unknown samples.

• Validation: It is important to validate that the recombinant MSP standard behaves similarly to the endogenous MSP in your samples (parallelism/dilution linearity). Differences in post-translational modifications or folding between recombinant and endogenous proteins can affect antibody binding and quantification accuracy.

• Carrier Protein: If your recombinant MSP is supplied with a carrier protein (e.g., BSA), this may affect your assay, especially if the carrier interferes with detection. Carrier-free versions are preferred for ELISA standard curves.

• Concentration and Purity: Ensure the recombinant MSP is of high purity (>80% by SDS-PAGE) and that its concentration is accurately determined (e.g., by amino acid analysis or HPLC), not just by the manufacturer’s label.

Recommendations:

• Use the recombinant MSP standard in the same buffer and matrix as your samples to minimize matrix effects.
• Always include a standard curve in each ELISA run for accurate quantification.
• If possible, validate the standard with your specific ELISA kit or antibody pair.

In summary, Recombinant Human MSP can be used as a standard for ELISA quantification, but its suitability and accuracy depend on proper validation and compatibility with your assay system.

Recombinant Human MSP (Macrophage Stimulating Protein, also known as MST1) has been validated in published research primarily for applications involving activation of the MSP receptor (RON/MST1R) in cell-based assays, especially in studies of cancer cell signaling and migration.

Key validated applications include:

• Cell signaling assays: Recombinant human MSP has been used to activate the RON receptor in human cancer cell lines (e.g., MDA-MB-453 breast cancer cells) to study downstream signaling pathways, such as phosphorylation of RON and subsequent cellular responses.
• Cell migration and invasion assays: MSP is frequently applied to assess its effect on cell motility, particularly in cancer research, where it is used to stimulate migration and invasion in vitro.
• ELISA standard: Recombinant MSP is used as a standard in enzyme-linked immunosorbent assays (ELISA) to quantify MSP levels in biological samples.
• Receptor-ligand binding studies: It is employed to characterize binding kinetics and specificity between MSP and its receptor RON/MST1R.

Additional context:

• MSP is a soluble cytokine in the plasminogen-related growth factor family, and its biological effects are mediated through the RON receptor, influencing processes such as cell proliferation, migration, and survival.
• While most published research focuses on cancer biology, MSP is also studied in the context of inflammation and tissue repair due to its role in macrophage activation.

No evidence was found in the provided search results for use in animal models or clinical studies; applications are primarily in vitro and cell-based. If you require information on a specific assay or disease context, please specify for a more targeted summary.

To reconstitute and prepare Recombinant Human MSP (Macrophage Stimulating Protein) for cell culture experiments, dissolve the lyophilized protein in sterile buffer—typically phosphate-buffered saline (PBS)—to a working concentration, ensuring gentle handling to preserve bioactivity.

Step-by-step protocol:

• Centrifuge the vial briefly to collect all lyophilized powder at the bottom.
• Add sterile PBS (or the buffer specified in the product datasheet) to achieve a final concentration of 100 μg/mL or as required for your assay. For example, add 100 μL PBS to 100 μg protein.
• Gently mix by pipetting or slow vortexing. Avoid vigorous shaking or foaming, which can denature the protein.
• Incubate at room temperature for 15–30 minutes to ensure complete dissolution.
• Aliquot the solution if not using immediately, and store at 2–8 °C for up to 1 month or –20 to –70 °C for longer-term storage. Avoid repeated freeze-thaw cycles to maintain protein integrity.
• Dilute further in cell culture medium or assay buffer as needed for your experiment.

Additional notes:

• If the protein was supplied with a carrier (e.g., BSA), this helps stabilize the protein during reconstitution and storage.
• Always consult the product datasheet for specific buffer recommendations, as some recombinant proteins may require additives (e.g., reducing agents, carrier proteins) for optimal stability.
• For cell culture, ensure all buffers and solutions are sterile and endotoxin-free to prevent confounding cellular responses.

Best practices:

• Use gentle agitation and avoid foaming.
• Prepare fresh working solutions for each experiment when possible.
• Store aliquots at recommended temperatures and avoid repeated freeze-thaw cycles.

This protocol ensures that Recombinant Human MSP is properly reconstituted and ready for reliable use in cell culture assays.