Recombinant Mouse IL-21R

Recombinant Mouse IL-21R is a valuable tool for research applications focused on immune regulation, cytokine signaling, and disease modeling, particularly in studies involving T cell, B cell, and macrophage function in mice.

Key scientific reasons to use recombinant mouse IL-21R include:

• Dissecting IL-21/IL-21R Signaling Pathways: IL-21R is the receptor for interleukin-21 (IL-21), a cytokine that modulates both innate and adaptive immune responses. Recombinant IL-21R enables precise investigation of how IL-21 influences immune cell differentiation, activation, and cytokine production.

• Functional Blocking and Bioassays: Recombinant IL-21R can be used as a decoy receptor or in Fc-chimera formats to inhibit IL-21-dependent cellular responses, such as IFN-γ secretion in NK cells, allowing for functional studies and mechanistic dissection of IL-21-mediated effects.

• Modeling Disease Mechanisms: IL-21/IL-21R signaling is implicated in various disease models, including infection, autoimmunity, allergy, and cardiovascular pathology. For example:

  • Promotes pro-inflammatory macrophage polarization and cytokine production during respiratory infection.
  • Modulates T cell-B cell interactions and humoral immunity, relevant for malaria and autoimmune disease models.
  • Influences myocardial hypertrophy and fibrosis in cardiac injury models.
  • Essential for epicutaneous sensitization and allergic skin inflammation.

• Cellular and Molecular Applications: Recombinant IL-21R is used in:

  • Flow cytometry to study receptor expression and ligand binding.
  • ELISA for quantifying IL-21 or IL-21R interactions.
  • Bioassays to measure functional outcomes of IL-21 signaling.
  • In vivo studies to assess the physiological role of IL-21R in mouse models.

• Therapeutic and Translational Research: Engineered IL-21R constructs are being explored to enhance T cell stemness, proliferation, and antitumor efficacy, as well as to prevent T cell exhaustion in immunotherapy contexts.

• Immunological Homeostasis and Differentiation: IL-21R is critical for regulating T and B cell proliferation, Ig class switching, and differentiation of effector and memory T cells.

In summary, using recombinant mouse IL-21R allows for controlled, reproducible studies of IL-21-mediated immune processes, supports mechanistic and therapeutic research, and provides essential reagents for cellular, molecular, and in vivo assays in mouse models.

Recombinant Mouse IL-21R can be used as a standard for quantification or calibration in ELISA assays only if your assay is specifically designed to measure IL-21R protein, not IL-21 cytokine.

Most ELISA assays for mouse IL-21 are designed to quantify the cytokine IL-21, not the IL-21 receptor (IL-21R). In these assays, the standard must be recombinant mouse IL-21, as the antibodies used are specific for IL-21 and will not recognize IL-21R. Using recombinant IL-21R as a standard in an IL-21 ELISA would result in inaccurate quantification because the assay is not calibrated for the receptor protein.

However, if you are performing an ELISA specifically for mouse IL-21R (for example, to quantify soluble IL-21R in biological samples), then recombinant mouse IL-21R is appropriate and commonly used as a standard. ELISA kits designed for IL-21R quantification use antibody pairs that specifically recognize IL-21R, and the standard provided or recommended is recombinant IL-21R.

Key considerations:

• Match the standard to the target analyte: Use recombinant mouse IL-21R only in assays designed to detect IL-21R, not IL-21.
• Check assay validation: Ensure your ELISA protocol or kit is validated for quantification of IL-21R using recombinant IL-21R as a standard.
• Formulation matters: For use as an ELISA standard, recombinant proteins are often supplied with carrier proteins (e.g., BSA) to improve stability and reproducibility.

Summary Table:

If your goal is to quantify IL-21R in your samples, recombinant mouse IL-21R is suitable as a standard. If you are quantifying IL-21, you must use recombinant mouse IL-21 as your standard. Always confirm the specificity and validation of your ELISA system before selecting a standard.

Recombinant Mouse IL-21R has been validated for several key applications in published research, primarily in immunological and cell signaling studies. The most frequently reported applications include:

• Bioassay: Used to assess IL-21-dependent biological activities, such as inhibition of IFN-γ secretion in NK-92 cells and modulation of immune cell function.
• Flow Cytometry: Applied to analyze cell surface expression and immune cell phenotyping, particularly in studies of T cell and B cell interactions, autoimmune responses, and humoral immunity.
• ELISA: Utilized for quantifying cytokine production and immune responses, especially in the context of T cell-B cell interactions and autoimmune disease models.
• Western Blot: Used to detect IL-21R protein expression and downstream signaling molecules in various cell types.
• Functional Assays (FuncS): Employed to validate receptor activity and ligand binding, confirming the biological relevance of the recombinant protein.

Supporting details and context:

• Bioassay validation is central for assessing the ability of recombinant IL-21R to inhibit IL-21-mediated effects, such as IFN-γ secretion in NK cells, and for studying immune modulation in vitro.
• Flow cytometry has been used to investigate the role of IL-21R in T cell differentiation, B cell function, and immune cell accumulation in lymphoid organs, including in models of malaria and autoimmune disease.
• ELISA is commonly used to measure cytokine levels (e.g., IL-4, IL-13, IFN-γ) in splenocyte cultures and to assess humoral immune responses following antigen sensitization.
• Western blot applications include detection of IL-21R and related signaling proteins in immune cells, supporting mechanistic studies of cytokine signaling.
• Functional assays (including SDS-PAGE and ligand binding studies) confirm the activity and purity of the recombinant protein, ensuring its suitability for downstream applications.

Additional relevant information:

• Recombinant Mouse IL-21R has also been used in studies of allergic skin inflammation, cardiac fibrosis, and infectious disease models to elucidate its role in immune regulation and tissue pathology.
• The protein is typically validated in whole cell systems, protein preparations, and in vivo models, reflecting its broad utility in immunological research.
• Published research highlights its importance in T cell-B cell interactions, Th17 and T follicular helper cell differentiation, and modulation of inflammatory responses.

In summary, Recombinant Mouse IL-21R is validated for bioassay, flow cytometry, ELISA, Western blot, and functional assays, with broad applications in immunology, cell signaling, and disease modeling.

To reconstitute and prepare Recombinant Mouse IL-21R protein for cell culture experiments, follow these general steps, which are consistent with best practices for recombinant cytokine receptor proteins:

1. Centrifuge the vial before opening to ensure all lyophilized material is at the bottom.

2. Reconstitution:

   • Add sterile distilled water or sterile PBS to the vial. The recommended concentration for reconstitution is typically 0.1–0.5 mg/mL in sterile distilled water, or 100 μg/mL in sterile PBS if specified by the product datasheet.
   • Gently mix by pipetting up and down or swirling. Do not vortex, as this can denature the protein.

3. Carrier Protein (Optional but Recommended for Stability):

   • For long-term storage or if the protein will be diluted to low concentrations, add a carrier protein such as 0.1% BSA (bovine serum albumin) or HSA (human serum albumin) to prevent adsorption and increase stability.
   • If your application is sensitive to carrier proteins (e.g., certain binding assays), use the carrier-free formulation and avoid adding BSA/HSA.

4. Aliquoting and Storage:

   • After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
   • Store aliquots at –20°C or –70°C for long-term storage, or at 2–8°C for short-term use (up to 1 month).
   • Avoid repeated freeze-thaw cycles.

5. Preparation for Cell Culture:

   • Before adding to cell cultures, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or buffer.
   • If using a carrier protein, ensure the final concentration in your working solution does not interfere with your assay.

Summary Table: Recombinant Mouse IL-21R Reconstitution

Additional Notes:

• Always consult the specific product datasheet for any unique instructions, as formulations may vary.
• If the protein is supplied as a fusion (e.g., Fc chimera), the same general reconstitution principles apply.
• For functional assays, confirm activity with a pilot experiment, as some proteins may require gentle handling or specific buffers for optimal activity.

These guidelines are based on standard protocols for recombinant mouse IL-21R and similar cytokine receptors.