Recombinant Mouse CXCL16

Recombinant Mouse CXCL16 is widely used in research applications to study immune cell recruitment, inflammation, tissue regeneration, and tumor immunology due to its well-characterized bioactivity and reproducibility in experimental systems.

Key scientific reasons to use recombinant Mouse CXCL16:

• Immune Cell Recruitment: CXCL16 is a chemokine that binds to the CXCR6 receptor, mediating the recruitment of lymphocyte subsets (including T cells, macrophages, and neutrophils) during inflammation and immune responses. This makes it valuable for dissecting mechanisms of immune cell trafficking in vitro and in vivo.

• Muscle Regeneration and Fibrosis: CXCL16 is critical for muscle regeneration by stimulating satellite cell proliferation and differentiation, and by regulating macrophage recruitment to injured tissue. CXCL16 knockout models show impaired muscle regeneration and increased fibrosis, highlighting its essential role in tissue repair.

• Tumor Immunology: CXCL16 promotes the recruitment of activated CD8^+^ T cells into tumors, enhancing antitumor immunity and synergizing with other chemokines to augment immune responses against cancer. Recombinant CXCL16 can be used to model or manipulate tumor microenvironments in preclinical studies.

• Inflammatory and Cardiovascular Research: CXCL16 is upregulated in heart failure and can stimulate matrix metalloproteinase (MMP) activity in cardiomyocytes and fibroblasts, influencing tissue remodeling and inflammation.

• Th1 Immune Responses: CXCL16 promotes IFN-γ production by NKT cells and supports Th1-skewed immune responses, making it useful for studies on cytokine signaling and immune polarization.

Technical advantages of recombinant protein:

• Defined Activity and Purity: Recombinant Mouse CXCL16 is typically >97% pure and validated for bioactivity, ensuring reproducible results in cell-based assays and mechanistic studies.
• Controlled Experimental Conditions: Using recombinant protein allows precise control over concentration, timing, and context of CXCL16 exposure, facilitating mechanistic dissection of its effects.
• Species-Specificity: Mouse CXCL16 shares high sequence identity with human CXCL16 in the chemokine domain, but recombinant mouse protein is essential for murine models to ensure physiological relevance.

Common applications include:

• Chemotaxis assays to study immune cell migration.
• In vitro stimulation of primary cells (e.g., satellite cells, macrophages, T cells).
• In vivo administration in mouse models of inflammation, tissue injury, or cancer.
• Mechanistic studies of CXCL16/CXCR6 signaling pathways.

Summary:
Use recombinant Mouse CXCL16 in your research to precisely investigate its roles in immune cell recruitment, tissue regeneration, inflammation, and tumor immunity, leveraging its validated bioactivity and reproducibility for robust experimental outcomes.

Yes, recombinant Mouse CXCL16 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for your specific assay system. Recombinant proteins are commonly used as standards in quantitative ELISA protocols to generate standard curves for accurate measurement of analyte concentrations in biological samples.

Essential context and supporting details:

• ELISA Standard Curve: Quantitative ELISA kits for Mouse CXCL16 typically use either recombinant or natural CXCL16 as the standard. The recombinant protein is serially diluted to create a standard curve, which is then used to interpolate the concentration of CXCL16 in unknown samples.
• Expression System: The recombinant Mouse CXCL16 standard may be produced in E. coli, yeast, or mammalian cells. The expression system can affect post-translational modifications, which may influence antibody recognition and assay performance. It is important to confirm that the recombinant standard is compatible with the antibodies used in your ELISA.
• Validation: Before using recombinant Mouse CXCL16 as a standard, ensure it is validated for your assay. This includes confirming linearity, recovery, and parallelism with native CXCL16 in your sample matrix. Most commercial ELISA kits provide validation data showing that recombinant CXCL16 is suitable for use as a standard.
• Preparation: Follow the manufacturer’s instructions for reconstitution and dilution of the recombinant standard. For example, some protocols recommend reconstituting the standard in a specific diluent and performing serial dilutions to cover the assay’s dynamic range.
• Assay Range: Typical standard curve ranges for Mouse CXCL16 ELISA are from approximately 1–2000 pg/mL, depending on the kit and protocol.

Additional relevant information:

• Research Use: Recombinant Mouse CXCL16 standards are intended for research use only and not for diagnostic applications.
• Precision: Assay precision (intra- and inter-assay CVs) should be assessed to ensure reliable quantification when using recombinant standards.
• Documentation: Always document the lot number, concentration, and preparation method of the recombinant standard used for calibration to ensure reproducibility.

Summary Table: Use of Recombinant Mouse CXCL16 as ELISA Standard

In summary: Recombinant Mouse CXCL16 is suitable as a standard for ELISA quantification, provided it is validated for your assay and prepared according to protocol.

Recombinant Mouse CXCL16 has been validated primarily for bioactivity assays and functional studies in published research, with applications including immune cell recruitment, inflammation models, and tumor immunology.

Key validated applications in published research include:

• Bioactivity assays: Recombinant Mouse CXCL16 is routinely used to confirm its ability to bind and activate its receptor CXCR6, and to induce chemotaxis of lymphocyte subsets, especially CD8⁺ T cells.
• Functional assays: These include studies of immune cell migration, proliferation, and activation, particularly in the context of inflammation and cancer.
• ELISA (Enzyme-Linked Immunosorbent Assay): Used as a standard or control protein for quantifying CXCL16 levels in biological samples.
• Tumor immunology models: Recombinant CXCL16 has been used to investigate its role in recruiting T cells to tumor sites, enhancing antitumor immunity, and modulating the tumor microenvironment in mouse models of cancer.
• Inflammation and autoimmune disease models: CXCL16 is studied for its role in attracting lymphocytes during inflammatory responses and in diseases such as multiple sclerosis and diabetes.
• Cell signaling and receptor binding studies: Used to characterize the interaction between CXCL16 and CXCR6, and downstream signaling pathways.

Additional validated protocols include:

• SDS-PAGE and HPLC: For protein purity and integrity assessment.
• In vitro and in vivo migration assays: To measure chemotactic activity on specific immune cell populations.

In summary, Recombinant Mouse CXCL16 is validated for use in bioactivity and functional assays, ELISA, immune cell migration studies, and tumor immunology research, with supporting evidence from multiple published studies and product validations.

To reconstitute and prepare Recombinant Mouse CXCL16 protein for cell culture experiments, dissolve the lyophilized protein in sterile PBS or sterile water to a concentration of at least 100 μg/mL.

Step-by-step protocol:

• Equilibrate the vial to room temperature before opening to minimize condensation.
• Centrifuge the vial briefly to collect the lyophilized powder at the bottom.
• Add sterile PBS or sterile water directly to the vial to achieve a final concentration of 100 μg/mL or higher. For example, if you have 25 μg of protein, add 250 μL to reach 100 μg/mL.
• Gently mix by pipetting up and down or by gentle vortexing. Avoid vigorous agitation to prevent protein denaturation.
• Incubate at room temperature for 10–30 minutes to ensure complete dissolution.
• If required for your application, further dilute the reconstituted stock in cell culture medium or an appropriate buffer. For sensitive cells, consider adding 0.1% BSA as a carrier protein to stabilize the protein during dilution.
• Filter sterilize the working solution if sterility is a concern and the protein is not already sterile-filtered.

Storage and handling:

• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at –20°C to –70°C for long-term storage, or at 2–8°C for short-term use (up to 1 month).
• Avoid multiple freeze-thaw cycles to maintain protein integrity.

Additional notes:

• Endotoxin levels in commercial recombinant proteins are typically low, but if your application is highly sensitive, confirm the endotoxin specification (<1 EU/μg is standard).
• For cell culture experiments, always use sterile technique and reagents to prevent contamination.

Summary Table:

This protocol ensures the recombinant CXCL16 is ready for use in cell-based assays, chemotaxis, or other functional studies. Adjust concentration and buffer as needed for your specific experimental design.