Recombinant Human IL-2 Rβ

Recombinant Human IL-2 Rβ is used in research applications to specifically study and manipulate the IL-2 signaling pathway, which is central to T cell biology, immune modulation, and adoptive cell therapies. Its recombinant form enables controlled experimental conditions, high reproducibility, and the ability to dissect receptor-ligand interactions with precision.

Key scientific applications and advantages include:

• Mechanistic Studies: Recombinant IL-2 Rβ allows detailed investigation of how IL-2 interacts with its receptor, facilitating analysis of downstream signaling events, T cell proliferation, and differentiation.
• Cellular Expansion and Activation: When paired with IL-2 or engineered orthogonal IL-2 variants, recombinant IL-2 Rβ can be used to selectively expand and activate specific T cell populations, such as CD4+ or CAR T cells, without undesired activation of regulatory T cells (Tregs).
• Immunotherapy Research: In adoptive cell therapy models, expressing recombinant IL-2 Rβ on engineered T cells enables tunable control over cell expansion and persistence, potentially improving antitumor efficacy while minimizing systemic toxicity.
• Blocking and Functional Assays: Recombinant IL-2 Rβ is essential for antibody blocking experiments, receptor binding assays, and for validating the specificity of IL-2–mediated responses in vitro and in vivo.
• Orthogonal Cytokine Systems: Modified recombinant IL-2 Rβ can be used in orthogonal cytokine-receptor systems, which allow selective stimulation of engineered immune cells, enhancing safety and efficacy in preclinical models.

Best practices for using recombinant IL-2 Rβ include:

• Employing it in combination with recombinant IL-2 or its engineered variants to dissect receptor-specific effects.
• Utilizing it in dose-response and time-course experiments to quantify signaling dynamics.
• Applying it in blocking assays to confirm the specificity of IL-2–mediated biological functions.

In summary, recombinant human IL-2 Rβ is a powerful tool for immunological research, enabling precise modulation and analysis of IL-2–dependent cellular processes, with broad applications in basic science, translational research, and therapeutic development.

Recombinant Human IL-2 Rβ (Interleukin-2 Receptor Beta) is not suitable as a standard for quantification or calibration in ELISA assays designed to measure IL-2 cytokine levels. ELISA standards must match the analyte being quantified—in this case, human IL-2, not its receptor subunit.

Key points:

• ELISA standards for IL-2 quantification are typically recombinant human IL-2 protein, not the IL-2 receptor or its subunits.
• The IL-2 Rβ protein is a component of the IL-2 receptor complex and is structurally and functionally distinct from the IL-2 cytokine itself. Using IL-2 Rβ as a standard would not generate a valid standard curve for IL-2 quantification, as the antibodies in IL-2 ELISAs are specific for the IL-2 cytokine, not its receptor.
• Best practice is to use a highly purified or recombinant IL-2 protein as your standard, prepared and quantified according to established protocols.

If your goal is to quantify IL-2 receptor beta (IL-2 Rβ) itself (for example, soluble IL-2Rβ in biological fluids), you would need an ELISA specifically designed for IL-2 Rβ, with a corresponding IL-2 Rβ standard. For quantifying IL-2 cytokine, always use an IL-2 protein standard.

Summary Table:

Conclusion:
Do not use recombinant human IL-2 Rβ as a standard for IL-2 quantification ELISAs. Use a recombinant human IL-2 protein standard that matches your assay’s target and antibody specificity.

Recombinant Human IL-2 Rβ (Interleukin-2 Receptor Beta) has been validated in published research primarily for applications involving immune cell signaling, T cell proliferation, and as a component in engineered cytokine-receptor systems for immunotherapy.

Key validated applications include:

• Functional studies of IL-2/IL-2Rβ signaling: Recombinant IL-2Rβ has been used to characterize the interaction between IL-2 and its receptor, demonstrating its role in enhancing the proliferation of CD4+ T lymphocytes and modulating immune responses. These studies often use recombinant IL-2Rβ in binding assays (such as pull-down and ELISA) and functional assays to assess T cell activation and proliferation.

• Engineering of orthogonal cytokine-receptor systems: Recombinant human IL-2Rβ has been engineered with specific mutations to create orthogonal (non-cross-reactive) receptor-cytokine pairs. These systems have been validated in vitro and in vivo to selectively stimulate engineered T cells (such as CAR T cells) without activating endogenous T cells, thereby enhancing antitumor efficacy and reducing systemic toxicity. Functional validation includes proliferation assays, cytokine signaling analysis, and in vivo expansion of engineered T cells in mouse models.

• Receptor binding and pharmacological profiling: Recombinant IL-2Rβ has been used in surface plasmon resonance (SPR) and other binding assays to quantify the affinity and kinetics of IL-2 or engineered IL-2 variants for the receptor. This is critical for the development of modified IL-2 therapeutics with altered receptor selectivity and improved pharmacological properties.

• Blocking and mechanistic studies: Antibodies against recombinant IL-2Rβ have been used to block IL-2 signaling in functional assays, confirming the essential role of IL-2Rβ in mediating IL-2-driven biological effects such as T cell proliferation and cytokine production.

• Immunophenotyping and cell subset analysis: Recombinant IL-2Rβ has been used to identify and quantify IL-2Rβ+ cell populations in various tissues, supporting studies on immune cell subset dynamics in health and disease.

Summary Table: Applications of Recombinant Human IL-2 Rβ in Published Research

Note: While some studies referenced here use recombinant IL-2Rβ from non-human species (e.g., flounder), the methodologies and validation approaches are directly applicable to human recombinant IL-2Rβ, and human-specific studies have validated similar applications, especially in the context of immunotherapy and engineered cell therapies.

To reconstitute and prepare Recombinant Human IL-2 Rβ protein (also known as CD122) for cell culture experiments, follow these general best practices, which are based on protocols for recombinant cytokine and receptor proteins:

1. Reconstitution:

• Buffer: Use sterile phosphate-buffered saline (PBS), pH 7.2–7.4, or sterile distilled water as the solvent, unless the product datasheet specifies otherwise. For enhanced stability and to prevent protein loss, include a carrier protein such as 0.1%–1% human serum albumin (HSA) or bovine serum albumin (BSA).
• Concentration: A typical reconstitution concentration is 0.1–0.2 mg/mL. Adjust the volume of buffer added to the lyophilized protein to achieve your desired stock concentration.
• Technique: Briefly centrifuge the vial to collect the powder at the bottom. Add the buffer gently along the vial wall to avoid foaming. Swirl or gently tap the vial to mix; do not vortex or shake vigorously.

2. Aliquoting and Storage:

• Aliquot: Divide the reconstituted protein into small aliquots to avoid repeated freeze-thaw cycles, which can degrade protein activity.
• Storage: Store aliquots at –20°C to –80°C for long-term storage. For short-term use (up to 1 week), store at 2–8°C.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

3. Working Solution Preparation:

• For cell culture, dilute the stock solution to the desired working concentration using cell culture medium or PBS containing a carrier protein (e.g., 0.1% BSA or HSA) to minimize adsorption to plasticware.
• Prepare working solutions fresh before use, or store at 2–8°C for up to one week if stability data supports this.

4. Sterility:

• Work under aseptic conditions to prevent contamination.
• If necessary, filter the final working solution through a 0.22 μm sterile filter. However, some protocols recommend not filtering the reconstituted protein itself if it is already sterile, to avoid protein loss.

5. Additional Notes:

• Always consult the specific product datasheet for any unique requirements, as recombinant proteins may have formulation-specific instructions.
• If the protein is provided in a carrier-free form, the addition of a carrier protein is especially important to prevent loss due to adsorption.

Example Protocol for Recombinant Human IL-2 Rβ (CD122):

1. Briefly centrifuge the vial.
2. Add sterile PBS (pH 7.2–7.4) containing 0.1% HSA to achieve 0.2 mg/mL.
3. Gently swirl to dissolve completely.
4. Aliquot and store at –80°C.
5. For cell culture, dilute aliquots in medium with 0.1% HSA or BSA.

Summary Table:

These guidelines are consistent with standard protocols for recombinant cytokine receptors and specifically for IL-2 Rβ (CD122). Always verify with the product-specific datasheet for any unique requirements.