Recombinant Mouse IL-21

Recombinant Mouse IL-21 is widely used in research because it is a potent immunomodulatory cytokine that regulates multiple aspects of both innate and adaptive immune responses, making it valuable for studies in immunology, oncology, and cell biology.

Key scientific reasons to use recombinant mouse IL-21 in research applications:

• T and B Cell Modulation: IL-21 enhances the proliferation and survival of CD40-primed B cells, promotes plasma cell differentiation, and induces immunoglobulin class switching to IgG. It also co-stimulates activation, proliferation, and survival of CD8+ T cells and NKT cells, and enhances the differentiation of effector and central memory T cells.

• NK Cell Activation: IL-21 increases the cytotoxic activity and IFN-γ production of natural killer (NK) cells, induces their terminal differentiation, and supports their functional maturation.

• Th17 and Tfh Cell Differentiation: IL-21 supports the differentiation of pro-inflammatory Th17 cells and follicular helper T (Tfh) cells, which are critical for adaptive immunity and antibody responses.

• Regulation of Immune Homeostasis: IL-21 regulates T cell and hematopoietic progenitor cell homeostasis, and can antagonize regulatory T cell (Treg) differentiation, thus influencing immune tolerance and autoimmunity.

• Cancer and Antiviral Research: IL-21 has demonstrated antitumor and antiviral effects in preclinical models by enhancing cytotoxic T lymphocyte (CTL) responses and promoting immune-mediated tumor rejection. It is being explored as a therapeutic agent and as an adjuvant in combination with other immunotherapies.

• Functional Assays and Cell Culture: Recombinant mouse IL-21 is optimized for use in cell culture, differentiation studies, and functional assays, providing a controlled and reproducible way to study its effects on immune cells.

• Mechanistic Studies: IL-21 activates key signaling pathways (Jak1/Jak3, Stat1, Stat3, Stat5), making it useful for dissecting cytokine signaling mechanisms and their downstream effects.

In summary, recombinant mouse IL-21 is a versatile tool for dissecting immune cell function, modeling disease, and developing new immunotherapies due to its broad and potent effects on multiple immune cell types and pathways.

Yes, Recombinant Mouse IL-21 can be used as a standard for quantification or calibration in ELISA assays, provided it is suitable for your specific ELISA kit and experimental requirements.

Key Points:

• Recombinant Mouse IL-21 (such as the product from R&D Systems/Bio-Techne, catalog #594-ML) is commonly used as a standard in ELISA assays to generate a standard curve for quantifying endogenous mouse IL-21 in samples.
• It is recommended to use the recombinant protein with BSA for ELISA standards, as BSA helps stabilize the protein and reduces non-specific binding.
• Some vendors (e.g., BioLegend) specifically caution that carrier-free recombinant proteins are not recommended for bioassays but may still be suitable for ELISA standards if properly validated.
• Always confirm compatibility with your ELISA kit manufacturer’s recommendations and ensure the recombinant protein is properly diluted in the same matrix as your samples to minimize matrix effects.

Best Practices:

• Use the recombinant protein to prepare a serial dilution series for your standard curve.
• Follow the kit protocol for standard preparation and dilution.
• Store the recombinant protein according to the manufacturer’s instructions to maintain activity.

In summary, yes, recombinant Mouse IL-21 is appropriate for use as an ELISA standard, but always verify compatibility with your specific kit and assay conditions.

Recombinant Mouse IL-21 has been validated in published research for several key applications, primarily in bioassays, in vivo studies, cell culture, and immunoassays such as ELISA and Western blot.

Validated Applications in Published Research:

• Bioassays:
  Used to assess cytokine activity, such as induction of proliferation in B9 cells, modulation of T and B cell responses, and functional maturation of NK cells.
  Examples include:

  • Induction of B cell proliferation and class switching.
  • Promotion of Th17 differentiation and suppression of regulatory T cell (Treg) generation.
  • Enhancement of cytolytic activity and IFN-γ production in NK and CD8+ T cells.

• In Vivo Studies:
  Applied in mouse models to study immune modulation, autoimmune disease, and cancer immunotherapy.

  • Investigating IL-21’s role in autoimmune diseases (e.g., type 1 diabetes, rheumatoid arthritis).
  • Evaluating antitumor responses and synergy with immune checkpoint blockade therapies.
  • Studying effects on skin inflammation and recruitment of immune cells.

• Cell Culture:
  Used to stimulate or differentiate immune cells (T cells, B cells, NK cells, dendritic cells) in vitro.

  • Functional studies of cytokine signaling and immune cell interactions.
  • Assessment of plasma cell differentiation and IgG production.

• Immunoassays:

  • ELISA: Measurement of cytokine levels and immune responses in biological samples.
  • Western Blot: Detection of IL-21-induced signaling proteins and downstream effectors.

Additional Validated Uses:

• Regulation of IgE and IgG class switching in B cells.
• Suppression of dendritic cell function and mast cell degranulation.
• Assessment of STAT3-dependent signaling pathways.
• Evaluation of myocardial fibrosis and hypertrophy in cardiac models.

Summary Table of Applications

Key Notes:

• Most studies use recombinant mouse IL-21 to dissect immune cell functions, disease mechanisms, and therapeutic interventions in mouse models.
• Validation is robust for bioassays and in vivo immunological research, with additional support for cell culture and immunoassay protocols.

If you require protocol details or specific experimental setups for any of these applications, please specify the context or cell type of interest.

To reconstitute and prepare Recombinant Mouse IL-21 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer (typically PBS or water) to a concentration of 0.1–0.5 mg/mL, optionally including 0.1–1% carrier protein (such as BSA or HSA) to enhance stability during storage and use.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile buffer:
  • For most applications, use sterile PBS (pH 7.2–7.4) or sterile distilled water.
  • Typical reconstitution concentration is 0.1 mg/mL (100 μg/mL), but you may prepare up to 0.5 mg/mL if higher concentrations are needed.
• Carrier protein:
  • For improved stability, especially for storage or low-concentration working solutions, add 0.1–1% BSA or HSA to the buffer.
• Dissolve gently:
  • Gently pipette the solution down the sides of the vial to dissolve the protein. Avoid vortexing or vigorous pipetting to prevent denaturation.
  • Allow the solution to sit at room temperature for several minutes to ensure complete dissolution.
• Aliquot and storage:
  • Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
  • Store aliquots at –20°C to –80°C for long-term storage; at 2–8°C for short-term use (up to 1 week).
• Working solution:
  • Dilute the stock solution to the desired working concentration in cell culture medium immediately before use.
  • Typical working concentrations for cell culture assays range from 3–100 ng/mL, depending on the cell type and experimental design.

Additional notes:

• If using for bioassays, ensure the recombinant protein is labeled and validated for such use, as ELISA standards may not be suitable for cell culture.
• Avoid repeated freeze-thaw cycles, as this can reduce protein activity and stability.
• Always consult the specific product’s Certificate of Analysis or datasheet for lot-specific instructions, as formulations and recommended buffers may vary.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of recombinant mouse IL-21 for cell culture applications.