Recombinant Mouse IL-6

Recombinant Mouse IL-6 is widely used in research because it is a multifunctional cytokine essential for studying immune regulation, inflammation, hematopoiesis, and disease models in mice.

Key scientific reasons to use recombinant Mouse IL-6 in your research applications:

• Immune Cell Differentiation and Function: IL-6 is critical for the differentiation of Th17 cells, memory B cells, plasma cells, and the reciprocal regulation of Th17 and Foxp3(+) Treg cells. It inhibits TGF-β-induced Foxp3 and induces RORγt, a Th17 lineage-specific transcription factor, making it indispensable for dissecting T cell lineage commitment and immune tolerance.

• Inflammatory and Acute Phase Responses: IL-6 regulates acute phase reactions, inflammatory responses, and antigen-specific immune responses, making it a key molecule for modeling inflammation and immune activation in vitro and in vivo.

• Hematopoiesis and Stem Cell Biology: IL-6 enhances hematopoietic stem cell proliferation and supports the expansion of progenitor cells, which is vital for studies on blood cell development and regenerative medicine.

• Cell Proliferation and Bioassays: Recombinant Mouse IL-6 stimulates cell proliferation in specific cell lines (e.g., T1165.85.2.1 mouse plasmacytoma), making it useful for bioassays and functional studies of cytokine signaling.

• Metabolic and Stress Responses: IL-6 acts as a stress hormone coordinating systemic immunometabolic reprogramming, including the regulation of glucose metabolism during acute stress, which is relevant for neuroimmunology and metabolic research.

• Disease Modeling: IL-6 is implicated in the pathogenesis of autoimmune diseases, cancer, and chronic inflammatory conditions. Recombinant Mouse IL-6 is used to model these diseases, investigate cytokine signaling pathways, and test therapeutic interventions.

• Standardization and Reproducibility: Using recombinant protein ensures consistent, defined activity and purity, which is crucial for reproducible experimental results and for standardizing protocols across studies.

Typical applications include:

• In vitro cell culture assays to study immune cell activation, differentiation, and cytokine signaling.
• In vivo mouse models to investigate the role of IL-6 in disease pathogenesis, immune regulation, and therapeutic targeting.
• Functional bioassays to measure cytokine activity and downstream effects on target cells.

In summary, recombinant Mouse IL-6 is an essential tool for immunology, inflammation, hematopoiesis, and disease research due to its broad biological activities and relevance to mouse models.

Yes, recombinant mouse IL-6 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly prepared and matched to your assay system.

Supporting details:

• Recombinant mouse IL-6 is widely accepted as an ELISA standard for quantifying mouse IL-6 levels in various sample types, including serum, plasma, and cell culture supernatants.
• Assay compatibility: Most commercial and custom ELISA kits for mouse IL-6 are validated to detect both natural and recombinant forms, and standard curves generated with recombinant IL-6 are parallel to those with native IL-6.
• Preparation: For accurate quantification, reconstitute and dilute the recombinant IL-6 according to your assay protocol. Carrier proteins (e.g., BSA) may be recommended at concentrations of 5–10 mg/mL to stabilize the standard and prevent loss of activity. Always use the diluent specified by your kit or protocol to avoid matrix effects.
• Calibration: Many ELISA kits calibrate their standards against international reference materials, such as the NIBSC non-WHO Reference Material (93/730), ensuring traceability and consistency.
• Concentration range: Standard curves typically cover a range from high picogram to low nanogram per milliliter concentrations (e.g., 2000–15 pg/mL or 500–0 pg/mL), depending on assay sensitivity.

Best practices:

• Confirm that your recombinant IL-6 is of high purity (≥95%) and low endotoxin content, as these factors can affect assay performance.
• Validate that your antibodies (capture and detection) recognize recombinant IL-6 equivalently to native IL-6, especially if using custom assay components.
• Include a standard curve on each plate for accurate quantification and to account for inter-assay variability.
• Store recombinant IL-6 according to manufacturer recommendations, typically at ≤–20°C, and avoid repeated freeze-thaw cycles.

Limitations:

• Use recombinant mouse IL-6 only for research purposes, not for diagnostic applications, unless specifically validated.
• Do not use mouse IL-6 standards for quantifying IL-6 from other species, as cross-reactivity may be limited.

Summary: Recombinant mouse IL-6 is suitable and commonly used as a standard for ELISA quantification, provided it is prepared and handled according to best practices and assay-specific protocols.

Recombinant Mouse IL-6 has been validated for a broad range of applications in published research, primarily in vitro and in vivo functional assays, cell culture, and immunological studies.

Key validated applications include:

• Cell proliferation assays: Used to stimulate proliferation of mouse B cell lines and plasmacytoma cell lines (e.g., T1165.85.2.1).
• Bioassays: Widely used in functional bioassays to assess cytokine activity, including studies on T cell differentiation (e.g., Th17, Th9, Tfh), dendritic cell function, and hepatocyte expansion.
• Colony formation assays: Induction of colony formation from hematopoietic progenitor cells in semi-solid medium.
• Hybridoma preparation: Replacement of feeder cells in the generation of murine hybridomas.
• In vitro differentiation: Used for differentiation of Th17 cells and other T cell subsets.
• Retroviral transduction: Cultivation of bone marrow cells for retroviral transduction protocols.
• ELISA standard: Used as a standard in ELISA assays for quantifying IL-6 levels.
• Metabolic studies: Employed in metabolic research to model stress-induced hyperglycemia and hepatic gluconeogenesis in mice.
• Osteoclastogenesis: Stimulation of osteoclast formation from precursor cells.
• Acute phase response studies: Investigation of IL-6’s role in the acute phase response and inflammation.
• Protein-protein interaction and binding assays: Used in binding and protein interaction studies.

Published research examples:

• Studies on T cell subset differentiation and cytokine signaling (e.g., Th17, Th9, Tfh cells).
• Investigation of IL-6’s role in tumor biology, chemoresistance, and immune modulation.
• Research on metabolic adaptation to stress and glucose homeostasis in murine models.
• Use as a standard in immunoassays such as ELISA for cytokine quantification.

Summary Table of Validated Applications

These applications are supported by both product validation data and peer-reviewed publications, demonstrating the versatility of recombinant mouse IL-6 in immunology, hematology, oncology, and metabolic research.

To reconstitute and prepare Recombinant Mouse IL-6 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer to a concentration of 0.1–0.5 mg/mL, typically using sterile PBS with 0.1–1% carrier protein (such as BSA or HSA) to enhance stability and prevent adsorption to surfaces.

Detailed protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile buffer: Reconstitute the protein at 0.1–0.5 mg/mL in sterile PBS or distilled water. For optimal stability, include at least 0.1%–1% BSA or HSA as a carrier protein.
• Mix gently: Swirl or invert the vial gently until the protein is fully dissolved. Avoid vigorous vortexing to prevent denaturation.
• Allow to equilibrate: Let the solution sit at room temperature for 5–10 minutes to ensure complete dissolution.
• Aliquot and store: Divide into single-use aliquots to avoid repeated freeze-thaw cycles. Store at –20 °C or –80 °C for long-term storage; short-term storage (up to 1 month) can be at 2–8 °C under sterile conditions.
• Working solution: Dilute the stock solution to the desired working concentration (typically in the range of 1–100 ng/mL for cell culture) using cell culture medium or buffer containing carrier protein.

Additional notes:

• If the protein is supplied carrier-free, adding carrier protein during reconstitution is highly recommended to prevent loss due to adsorption.
• Avoid repeated freeze-thaw cycles, as this can reduce protein activity.
• Always use sterile technique to prevent contamination.

Example reconstitution (for 100 μg vial):

Add 1 mL sterile PBS + 0.1% BSA to obtain a 100 μg/mL stock solution.

This stock can then be further diluted in cell culture medium as needed for your experiments.

Summary of best practices:

• Use sterile PBS or water with 0.1–1% BSA/HSA.
• Reconstitute at 0.1–0.5 mg/mL.
• Mix gently, aliquot, and store appropriately.
• Dilute to working concentration in medium with carrier protein for cell culture use.