The predicted molecular weight of Recombinant Mouse IL-18 is Mr 18.2 kDa.
Predicted Molecular Mass
18
Formulation
This recombinant protein was lyophilized from a 0.2 μm filtered solution in phosphate buffered saline (PBS), with 1mM EDTA and 1 mM DTT with 5% trehalose, pH 7.2.
Storage and Stability
The lyophilized protein should be stored desiccated at -20 to -70°C. The reconstituted protein can be stored for at least one month at 2 to 8°C. For long-term storage of the reconstituted protein, aliquot into working volumes and store at -20 to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.
Applications and Recommended Usage ? (Quality Tested by Leinco)
ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 5 µg/ml (100 µl/well) µg/ml. Flow Cytometry:PN:R1190 Western Blotting: This antibody can be used to detect Human, Mouse and Rat TIM-1 by Western blot analysis at a concentration of 1.0-2.0 µg/ml when used in conjunction with compatible secondary reagents, such as PN:R1190, under either reducing or non-reducing conditions. The positive control for Western blotting is PN:U124.
Leinco Protein Advisor
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Recombinant Mouse IL-18 is widely used in research because it is a potent pro-inflammatory cytokine that plays a central role in regulating both innate and adaptive immune responses, making it valuable for studying immune activation, inflammation, cancer immunotherapy, and autoimmune disease mechanisms.
Key reasons to use recombinant mouse IL-18 in research applications:
Immune Cell Activation: IL-18 stimulates the production of interferon-gamma (IFN-γ) in T-helper type 1 (Th1) cells and natural killer (NK) cells, enhancing their cytotoxic activity and supporting studies on immune cell function and cytokine signaling.
Modeling Inflammatory and Autoimmune Diseases: IL-18 is implicated in the pathogenesis of various inflammatory and autoimmune diseases, such as psoriasis, inflammatory bowel disease, and systemic lupus erythematosus, making it useful for disease modeling and therapeutic target validation.
Cancer Immunology: IL-18 enhances anti-tumor immunity by activating T and NK cells, promoting dendritic cell maturation, and modulating the tumor microenvironment. Recombinant IL-18 is used to investigate anti-tumor mechanisms and as a tool in preclinical immunotherapy studies.
Protein-Protein Interaction and Antibody Development: Recombinant IL-18 can serve as an antigen for generating and validating antibodies, and as a bait protein in pull-down assays to study receptor interactions and downstream signaling pathways.
Standardization and Reproducibility: Recombinant proteins provide consistent quality and defined activity, enabling reproducible results in bioassays, cell culture experiments, and in vivo studies.
Functional Assays: It is commonly used as a positive control in assays measuring IFN-γ production, immune cell activation, and dose-response relationships in primary mouse splenocytes or established cell lines.
Additional technical advantages:
High Purity and Tagging: Recombinant mouse IL-18 is typically supplied at high purity and may include affinity tags (e.g., His-tag) for easy purification and quantification, facilitating downstream applications.
Biological Activity: The recombinant form retains full-length structure and biological activity, ensuring relevance for in vitro and in vivo studies.
In summary, recombinant mouse IL-18 is a versatile tool for dissecting immune mechanisms, modeling disease, developing immunotherapies, and standardizing immunological assays in mouse systems.
Yes, recombinant Mouse IL-18 can be used as a standard for quantification or calibration in ELISA assays, provided it is of appropriate purity and formulation. Recombinant cytokines like Mouse IL-18 are commonly used as standards to generate calibration curves for quantitative ELISA, allowing accurate determination of IL-18 concentrations in biological samples.
Key considerations for use as an ELISA standard:
Formulation: Recombinant Mouse IL-18 is often supplied either carrier-free or with BSA (bovine serum albumin) as a stabilizer. For ELISA standard preparation, the BSA-containing formulation is generally recommended to improve protein stability and minimize adsorption to plasticware.
Reconstitution: Follow the manufacturer’s instructions for reconstitution, typically in PBS or assay diluent, to achieve the desired concentration (e.g., 100 μg/mL).
Purity and Activity: Ensure the recombinant protein is highly purified and bioactive, as confirmed by SDS-PAGE and functional assays (e.g., induction of IFN-γ secretion in mouse T cells).
Compatibility: Most commercial Mouse IL-18 ELISA kits are validated to detect both natural and recombinant Mouse IL-18, with no significant cross-reactivity with other cytokines. Always verify that your ELISA kit recognizes recombinant Mouse IL-18 as a standard.
Standard Curve Preparation: Prepare serial dilutions of recombinant Mouse IL-18 in the same buffer or diluent used for your samples to minimize matrix effects and ensure accurate quantification.
Best Practices:
Use double determinations (duplicates) for each standard point to ensure reliability.
Avoid repeated freeze-thaw cycles of the recombinant standard to maintain protein integrity.
Include internal controls to confirm assay accuracy and consistency across runs.
Summary Table: Recombinant Mouse IL-18 as ELISA Standard
In conclusion, recombinant Mouse IL-18 is suitable as a standard for ELISA quantification, provided you follow best practices for preparation and validation within your specific assay system.
Recombinant Mouse IL-18 has been validated for multiple applications in published research, primarily in bioassays, cell culture, and in vivo studies to investigate immune modulation, cytokine signaling, and disease models.
Key validated applications include:
Bioassays: Used to induce IFN-γ secretion by activated mouse T cells, demonstrating its potency in stimulating immune responses. IL-18 bioactivity is commonly measured by its ability to trigger cytokine production in various cell types.
Cell Culture: Applied to whole cells and organoids to study immune cell activation, dendritic cell maturation, and epithelial responses. For example, IL-18 has been used to promote myeloid dendritic cell maturation and to trigger neutrophil respiratory burst.
In Vivo Studies: Extensively used in mouse models to assess its role in inflammation, autoimmune diseases, cancer immunotherapy, and tissue injury. Published studies have validated recombinant mouse IL-18 for:
Enhancing anti-tumor immunity and modulating tumor microenvironment.
Accelerating cardiac inflammation and dysfunction during ischemia/reperfusion injury.
Modulating neuronal activity and suppressing aberrant transmission in neurodegenerative disease models.
Investigating autoimmune and inflammatory disease mechanisms, such as in models of diabetes, psoriasis, and inflammatory bowel disease.
ELISA Standard: Recombinant mouse IL-18 is validated as a standard for quantifying IL-18 levels in immunoassays.
Organoid Models: Used to study epithelial defense circuits and cytokine-mediated tissue responses.
Additional validated research areas include:
Cancer immunotherapy: Engineered IL-18 variants have been tested for improved stability and efficacy in activating T and NK cells, both alone and in combination with checkpoint inhibitors.
Autoimmunity and inflammation: IL-18 has been implicated in the pathogenesis and regulation of autoimmune diseases, metabolic syndromes, and tissue-specific inflammatory responses.
In summary, recombinant mouse IL-18 is a well-established tool for:
In vivo models (disease pathogenesis, therapeutic intervention)
ELISA standards (quantification of IL-18)
These applications are supported by numerous peer-reviewed studies across immunology, oncology, neuroscience, and inflammation research.
To reconstitute and prepare Recombinant Mouse IL-18 protein for cell culture experiments, briefly centrifuge the vial to collect the contents, then add ice-cold, sterile distilled water (or PBS, depending on the formulation) to achieve the desired concentration, typically 100 μg/mL or as specified by the manufacturer. Perform all steps on ice to preserve protein stability.
Detailed protocol:
Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
Reconstitution volume: Commonly, add 250 μL of ice-cold, sterile distilled water for a 25 μg vial, or adjust proportionally based on your vial size and desired stock concentration (e.g., 100 μg/mL is standard for many applications).
Mix gently: Swirl or pipette gently to dissolve. Avoid vigorous vortexing to prevent protein denaturation.
Incubate on ice for several minutes to ensure complete dissolution.
Aliquot immediately after reconstitution to avoid repeated freeze-thaw cycles.
Storage after reconstitution:
Short-term (up to 1 week): 2–8 °C.
Long-term: Store aliquots at –80 °C.
Working dilution: Dilute the stock solution into your cell culture medium just before use. The optimal working concentration should be empirically determined for your specific assay, but typical ranges are 0.1–100 ng/mL depending on cell type and experimental design.
Additional notes:
If the protein is supplied with stabilizers (e.g., BSA, sucrose), reconstitute as directed in the product datasheet.
For carrier-free formulations, use only sterile water or PBS as recommended.
Avoid repeated freeze-thaw cycles, as this can reduce protein activity.
Always use low-retention tubes for aliquoting to minimize protein loss.
Example protocol (for a 25 μg vial):
1. Briefly centrifuge the vial.2. Add 250 μL ice-cold, sterile distilled water.3. Gently pipette to dissolve.4. Incubate on ice for 5–10 minutes.5. Aliquot and store at –80 °C.6. Dilute to working concentration in cell culture medium immediately before use.
Summary of key points:
Use ice-cold, sterile water or PBS for reconstitution.
Prepare aliquots and store at –80 °C.
Determine optimal working concentration empirically for your assay.
Always consult the specific datasheet for your recombinant IL-18 preparation, as formulations and recommended protocols may vary slightly between suppliers.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
