Recombinant Mouse uPAR

Recombinant Mouse uPAR is a valuable tool for research applications focused on cell migration, tissue remodeling, immune response, and disease modeling, particularly in cancer, inflammation, and wound healing studies.

Key scientific reasons to use recombinant mouse uPAR:

• Mechanistic Studies: uPAR is central to the regulation of proteolytic activity at the cell surface, primarily by binding urokinase-type plasminogen activator (uPA) and facilitating the conversion of plasminogen to plasmin. This process activates matrix metalloproteinases, enabling controlled extracellular matrix (ECM) degradation, cell migration, and tissue remodeling.

• Disease Modeling: Mouse uPAR is highly relevant for in vivo studies using murine models, allowing investigation of its role in cancer metastasis, immune cell recruitment, angiogenesis, and fibrinolysis. Recombinant forms enable precise manipulation of uPAR levels or function in genetically defined backgrounds.

• Therapeutic Target Validation: Recombinant mouse uPAR is used to test and validate therapeutic strategies, such as monoclonal antibodies, CAR-T cells, or small molecule inhibitors targeting uPAR, especially in preclinical cancer and inflammation models.

• Biomarker Research: uPAR expression correlates with disease progression, therapy resistance, and prognosis in various pathologies. Recombinant protein enables development and validation of diagnostic assays, including ELISA and immunoblotting, for quantifying uPAR in biological samples.

• Functional Assays: Recombinant mouse uPAR supports in vitro assays for cell adhesion, migration, invasion, and signaling pathway activation. It can be used to study interactions with uPA, integrins, and other cell surface receptors, dissecting downstream effects on PI3K/AKT, MAPK/ERK, and FAK/Src signaling.

• Immunological Studies: uPAR modulates immune cell recruitment and chemotaxis, making recombinant protein useful for investigating mechanisms of immune surveillance, inflammation, and vascular homeostasis in mouse models.

Best practices for use:

• Select recombinant mouse uPAR that matches the desired isoform (full-length, soluble, Fc-chimera) and is produced in a system ensuring proper folding and post-translational modifications.
• Use in combination with functional assays (migration, invasion, adhesion) and molecular techniques (immunoblotting, ELISA, FACS) to quantify and characterize uPAR activity and interactions.
• Employ in genetically modified mouse strains (e.g., uPAR knockout or overexpression) to dissect physiological and pathological roles.

Summary:
Using recombinant mouse uPAR enables precise, mechanistic, and translational research into cell migration, tissue remodeling, immune function, and disease progression, especially in murine models relevant to cancer, inflammation, and wound healing.

Yes, recombinant Mouse uPAR can be used as a standard for quantification or calibration in ELISA assays, provided it is well-characterized and compatible with your assay system. Recombinant uPAR is commonly used as a calibrator in commercial mouse uPAR ELISA kits, where it is supplied as a lyophilized standard to generate the standard curve for quantification.

Key considerations:

• Source and Characterization: The recombinant uPAR should be highly purified and its concentration accurately determined. Many kits use recombinant mouse uPAR expressed in mammalian cells (e.g., NS0 cells) to ensure proper folding and post-translational modifications.
• Assay Compatibility: The recombinant standard must be recognized by the capture and detection antibodies in your ELISA. Most commercial kits are validated with their supplied recombinant standard, so if you are using a different recombinant uPAR, confirm that it is immunologically equivalent to the native protein and compatible with your antibodies.
• Standard Curve Preparation: Follow best practices for preparing the standard curve, including serial dilutions in the same matrix as your samples (e.g., sample diluent or buffer), to minimize matrix effects and ensure accurate quantification.
• Documentation: Prepare the standard solution fresh as recommended (often within 2 hours of use) and store aliquots appropriately to maintain stability and activity.

Limitations and Best Practices:

• If your recombinant uPAR differs in glycosylation or structure from the endogenous protein, this may affect antibody recognition and quantification accuracy. Always validate the standard in your specific assay context.
• For highest accuracy, use a recombinant standard that matches the one used to calibrate your ELISA kit, or validate your recombinant standard against a known reference.
• Include internal controls and, if possible, spike recovery experiments to confirm that your standard behaves similarly to endogenous uPAR in your sample matrix.

Summary Table: Use of Recombinant Mouse uPAR as ELISA Standard

In conclusion, recombinant Mouse uPAR is suitable as a standard for ELISA quantification if it is well-characterized, compatible with your assay antibodies, and validated in your specific assay system.

Recombinant Mouse uPAR has been validated for a range of applications in published research, primarily in studies of cell signaling, cancer, inflammation, and tissue remodeling. The most commonly validated applications include:

• Western Blot: Used to detect uPAR expression in cell lysates, tissue homogenates, and recombinant protein preparations in mouse models of disease, including ischemic stroke, cancer, and inflammatory conditions.
• Flow Cytometry: Applied to analyze uPAR expression on the surface of mouse cells, including immune and tumor cells, and to study cell populations in cancer and immunology research.
• Immunohistochemistry/Immunofluorescence: Used to localize uPAR in mouse tissue sections, such as kidney and tumor tissues, to study spatial expression patterns.
• ELISA (Enzyme-Linked Immunosorbent Assay): Utilized for quantifying uPAR or its ligands in biological samples and for functional blocking assays to study receptor-ligand interactions.
• Mass Spectrometry: Employed for proteomic analysis of uPAR and its interacting partners in mouse models.
• Bioassays/Functional Assays: Used to assess the biological activity of uPAR, including its role in cell adhesion, migration, chemotaxis, and signal transduction.
• Blocking/Neutralization Studies: Recombinant mouse uPAR is used as a target in assays to test the efficacy of antibodies or small molecules in blocking uPA-uPAR interactions, relevant for therapeutic development.

Key research contexts where recombinant mouse uPAR has been validated:

• Cancer research: Studying tumor progression, metastasis, and the tumor microenvironment, including antibody-drug conjugate targeting and stromal interactions.
• Inflammation and immune response: Investigating uPAR’s role in inflammatory bowel disease, immune cell migration, and response to pathogens.
• Neurological disease: Assessing uPAR’s contribution to brain injury and neuroinflammation in mouse models of stroke.
• Cardiovascular and tissue remodeling: Exploring uPAR’s involvement in tissue repair, fibrosis, and vascular biology.

Summary Table: Validated Applications for Recombinant Mouse uPAR

These applications are supported by multiple peer-reviewed studies and product validation reports, demonstrating the versatility of recombinant mouse uPAR in both basic and translational research.

To reconstitute and prepare Recombinant Mouse uPAR protein for cell culture experiments, use sterile technique and follow these steps:

1. Centrifuge the vial briefly before opening to collect all lyophilized powder at the bottom.
2. Reconstitute the protein using sterile, distilled water or the buffer specified in the product datasheet (commonly PBS, pH 7.2, sometimes with 5% trehalose for stability). Gently add the liquid to the vial and avoid vigorous mixing to prevent foaming and protein denaturation.
3. Allow the protein to dissolve for 15–30 minutes at room temperature with gentle agitation.
4. Dilute to working concentration using sterile cell culture medium or buffer appropriate for your experiment. If the protein will be added directly to cell cultures, ensure the final buffer is compatible with your cells (e.g., serum-free medium if required).
5. For long-term storage, aliquot the reconstituted protein and store at −20°C to −80°C. Addition of a carrier protein such as 0.1% BSA or 5–10 mg/mL purified BSA is recommended to prevent adsorption and degradation, unless animal-derived proteins must be avoided (then use trehalose).
6. Avoid repeated freeze-thaw cycles by aliquoting into single-use portions.

Additional notes for cell culture:

• If using in serum-free conditions or in vivo, avoid carrier proteins like BSA or FBS; use trehalose as a stabilizer instead.
• Confirm the final concentration and buffer composition are suitable for your cell type and experimental design.
• Always consult the Certificate of Analysis (CoA) or datasheet for specific instructions regarding your recombinant protein batch.

Example protocol for reconstitution:

- Add sterile water or PBS (as specified) to the lyophilized protein to achieve the desired concentration.- Gently mix and let stand at room temperature for 15–30 minutes.- Dilute with cell culture medium or buffer to working concentration.- Aliquot and store unused portions at −20°C to −80°C with carrier protein if long-term storage is needed.

Key considerations:

• Use sterile technique throughout.
• Avoid vigorous mixing.
• Use carrier protein or trehalose for stability if storing aliquots.
• Check compatibility of buffer and additives with your cell culture system.

These steps ensure protein integrity and experimental reproducibility for cell culture applications.