Recombinant Rat IL-2

Recombinant Rat IL-2 is widely used in research because it is a key cytokine for modulating immune responses, particularly in rat models. It is essential for the proliferation and activation of T cells, the generation of cytotoxic T lymphocytes (CTLs), the stimulation of B cell proliferation and antibody production, and the maintenance of natural killer (NK) cells.

Key scientific reasons to use recombinant rat IL-2 in research applications:

• T Cell Proliferation and Activation: IL-2 is critical for the expansion of activated T cells, making it indispensable for studies involving T cell biology, immune activation, and adoptive cell transfer protocols.
• Regulatory T Cell (Treg) Expansion: IL-2 plays a central role in the expansion and maintenance of regulatory T cells, which are crucial for immune tolerance and the suppression of autoimmunity.
• Cytotoxic Lymphocyte Generation: It induces the generation of CTLs and supports NK cell persistence, both of which are important for anti-tumor and anti-viral immunity.
• B Cell Support: IL-2 facilitates B cell proliferation and immunoglobulin synthesis, supporting studies on humoral immunity.
• Autoimmunity and T Cell Homeostasis: IL-2 helps maintain T cell homeostasis and can suppress the development of Th17 cells, which are implicated in autoimmune diseases.
• Bioassay and Functional Studies: Recombinant rat IL-2 is used in bioassays to stimulate rat lymphocytes in vitro, enabling functional studies of immune cell responses, cytokine signaling, and immunomodulation.
• Cancer and Infectious Disease Models: IL-2 is used in preclinical models to study cancer immunotherapy, infectious disease resistance, and immune modulation, as it can enhance anti-tumor responses and modulate immune cell populations.
• Consistency and Reproducibility: Recombinant proteins provide high purity and batch-to-batch consistency, which are critical for reproducible experimental results.

Typical applications include:

• In vitro stimulation of rat T cells, NK cells, and B cells.
• Expansion of Tregs for studies of immune tolerance.
• Functional assays in immunology, oncology, and infectious disease research.
• Development of ELISA and other immunoassays for rat cytokine detection.

Summary:
Use recombinant rat IL-2 when you need a defined, species-specific cytokine to reliably stimulate, expand, or modulate rat immune cells in vitro or in vivo, especially for studies of T cell biology, immune regulation, cancer immunotherapy, and autoimmunity.

Yes, recombinant rat IL-2 can be used as a standard for quantification or calibration in ELISA assays, provided it matches the native protein in your samples and is prepared according to assay requirements.

Essential context and supporting details:

• ELISA Standard Use: Recombinant rat IL-2 is commonly used as a standard in ELISA kits designed to quantify rat IL-2 in biological samples such as serum, plasma, and cell culture supernatants. The standard curve generated with recombinant IL-2 allows for accurate quantification of IL-2 concentrations in unknown samples.

• Protein Form and Preparation: The recombinant protein should be of the same mature full-length sequence as native rat IL-2 to ensure assay compatibility. Carrier-free or BSA-containing formulations may be chosen depending on whether BSA interferes with your assay or sample matrix. It is important to prepare the standard dilutions freshly and follow the ELISA kit protocol for standard curve generation.

• Assay Validation: ELISA kits are validated to recognize both natural and recombinant rat IL-2, and dose-response curves for recombinant and native IL-2 are typically comparable. However, always confirm that your recombinant standard is compatible with the antibodies and detection system used in your specific ELISA.

• Best Practices:

  • Use the recombinant standard provided or recommended by your ELISA kit manufacturer when possible, as it is validated for that assay.
  • If using a recombinant standard from another source, ensure it is properly characterized (sequence, purity, activity) and matches the native protein in your samples.
  • Prepare standard dilutions according to the kit instructions and run a standard curve with each assay to ensure accurate quantification.
  • Store recombinant IL-2 according to manufacturer recommendations and include carrier protein (e.g., BSA) if required for stability.

• Limitations: If your recombinant IL-2 differs in sequence, glycosylation, or folding from the native protein, it may affect antibody recognition and quantification accuracy. Always verify equivalence with your assay system.

Summary of technical recommendations:

• Recombinant rat IL-2 is suitable as an ELISA standard if it matches the native protein and is prepared according to validated protocols.
• Follow your ELISA kit’s instructions for standard preparation and curve generation.
• Confirm compatibility between your recombinant standard and the antibodies used in your assay.

If you need protocol details for standard preparation or troubleshooting, please specify your ELISA format or detection system.

Recombinant Rat IL-2 has been validated for several applications in published research, primarily in bioassays, ELISA development, and functional studies involving immune cell modulation.

Key validated applications include:

• Bioassays: Recombinant rat IL-2 is widely used to assess its biological activity, such as stimulating T cell proliferation, modulating CD4+ T cell polarization, and supporting regulatory T cell expansion. Numerous studies have used it to probe immune cell function in vitro and in vivo, including:

  • Investigating T cell homeostasis and regulatory T cell maintenance.
  • Assessing the effects on Th17 cell development and autoimmunity suppression.
  • Studying lymphocyte migration and endothelial interactions.
  • Evaluating immune responses in disease models, such as renal fibrosis, autoimmune glomerulonephritis, and encephalomyelitis.
  • Enhancing diabetes development in rat models.

• ELISA Development: Recombinant rat IL-2 has been used as a standard or control in ELISA assays to quantify IL-2 levels in tissue homogenates and cell culture supernatants.

• Functional Assays: These include proliferation assays (e.g., using HT-2 cells), cytokine signaling studies, and immune cell differentiation protocols.

• Western Blot and Immunoprecipitation: While direct published examples for rat IL-2 are less common, recombinant IL-2 proteins (including rat) are validated for use in Western blotting and immunoprecipitation to detect IL-2 or confirm recombinant protein expression.

• Immunohistochemistry: Some recombinant IL-2 proteins are validated for immunohistochemical detection, though specific published examples for rat IL-2 are limited.

Summary Table: Validated Applications for Recombinant Rat IL-2

Notable Published Research Examples:

• Renal fibrosis attenuation via CD4+ T cell polarization in rats.
• Suppression of Th17 cells and amelioration of autoimmune glomerulonephritis.
• Vascular inflammation studies in systemic connective tissue disease models.
• Diabetes enhancement in BB rats.
• Autoimmune encephalomyelitis and innate immunity activation.

These applications demonstrate that recombinant rat IL-2 is a versatile tool for immunological research, especially for functional studies involving T cell biology, cytokine signaling, and disease modeling in rats.

To reconstitute and prepare Recombinant Rat IL-2 protein for cell culture experiments, follow these best-practice steps to ensure protein stability and biological activity:

1. Centrifuge the vial:
Briefly centrifuge the lyophilized protein vial before opening to ensure all material is at the bottom and to prevent loss of product when opening.

2. Reconstitution:

• Add sterile water or a suitable sterile buffer (such as PBS, pH 7.4) to the vial. The recommended concentration for stock solutions is typically 0.1–1.0 mg/mL.
• If the protein is carrier-free (no BSA), and you require additional stability, you may reconstitute in sterile buffer containing 0.1% BSA (cell culture grade) to help prevent adsorption and loss of activity.
• Gently pipette up and down to dissolve. Do not vortex as this can denature the protein.
• Allow several minutes for complete dissolution at room temperature.

3. Aliquoting and Storage:

• Once fully dissolved, aliquot the stock solution into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at –20°C or –80°C for long-term storage. Avoid repeated freeze-thaw cycles.

4. Working Solution Preparation:

• For cell culture, dilute the stock solution to the desired working concentration using sterile cell culture medium or buffer immediately before use.
• If further dilution is needed, use medium or buffer containing 0.1% BSA to maintain stability.

5. General Handling Tips:

• Use low-protein binding tubes for all steps to minimize loss.
• Avoid vigorous mixing or foaming.
• Use the reconstituted protein within 24 hours if stored at 2–8°C, or freeze immediately for longer storage.

Example Protocol:
Suppose you have 10 μg lyophilized IL-2:

• Add 100 μL sterile PBS with 0.1% BSA to achieve a 0.1 mg/mL (100 μg/mL) stock.
• Mix gently and let sit at room temperature for 10 minutes.
• Aliquot into 10 μL portions and freeze at –80°C.

Notes:

• Always consult the specific product datasheet for any manufacturer-specific recommendations regarding reconstitution buffer and concentration.
• For cell culture, ensure all reagents are endotoxin-free and sterile.

These steps will help preserve the biological activity and stability of recombinant rat IL-2 for reliable cell culture experiments.