Recombinant Mouse IL-3 Rβ

Recombinant Mouse IL-3 Rβ is used in research applications to study and manipulate the high-affinity receptor complex for interleukin-3 (IL-3), which is critical for hematopoietic cell proliferation, differentiation, and immune regulation.

The IL-3 receptor β subunit (IL-3 Rβ, also known as βc) is essential for forming the high-affinity IL-3 receptor complex. While the α subunit alone binds IL-3 with low affinity, the β subunit is required for high-affinity binding and signal transduction. The β subunit is also shared by the receptors for IL-5 and GM-CSF, making it a key molecule in cytokine signaling networks that regulate hematopoiesis and immune responses.

Key research applications for Recombinant Mouse IL-3 Rβ include:

• Cell signaling studies: Dissecting the molecular mechanisms of IL-3, IL-5, and GM-CSF signaling in hematopoietic and immune cells.
• Hematopoietic cell biology: Investigating the proliferation, differentiation, and survival of hematopoietic stem cells and lineage-committed progenitors, as the β subunit is required for IL-3-driven effects.
• Receptor binding assays: Characterizing receptor-ligand interactions, affinity, and specificity, especially in ELISA or sandwich immunoassays, where recombinant IL-3 Rβ can be used to detect or quantify receptor complexes.
• Functional studies: Exploring the role of βc in emergency hematopoiesis, inflammation, and immune cell recruitment, as it is a common signaling component for multiple cytokines involved in these processes.
• Disease modeling: Studying the impact of altered βc signaling in models of leukemia, autoimmune disease, or infection, given its role in myeloid lineage skewing and immune cell development.

Best practices:

• Use recombinant IL-3 Rβ in combination with IL-3 or other cytokines to reconstitute functional receptor complexes for in vitro assays.
• Employ recombinant IL-3 Rβ in ELISA or flow cytometry to quantify receptor expression or analyze receptor-ligand interactions.
• Store and handle recombinant proteins under sterile, low-temperature conditions to preserve bioactivity and stability.

In summary, Recombinant Mouse IL-3 Rβ is a critical tool for dissecting cytokine receptor biology, hematopoietic regulation, and immune signaling in mouse models, enabling precise manipulation and analysis of pathways relevant to health and disease.

Recombinant Mouse IL-3 Rβ should not be used as a standard for quantification or calibration in ELISA assays designed to measure IL-3 protein levels. ELISA standards must match the analyte being quantified—in this case, mouse IL-3, not its receptor subunit IL-3 Rβ.

Supporting details:

• ELISA assays for mouse IL-3 are calibrated using purified recombinant mouse IL-3 protein, not the receptor subunits. The standard curve must be generated with the same molecule as the target analyte to ensure accurate quantification.
• Recombinant Mouse IL-3 Rβ is a receptor protein, not the cytokine IL-3 itself. It is used in research for receptor binding studies or as a capture/detection reagent in immunoassays, but not as a quantification standard for IL-3.
• ELISA kits for mouse IL-3 specifically use recombinant mouse IL-3 as the standard, and their antibodies are raised against IL-3, not IL-3 Rβ.
• Using a receptor protein (IL-3 Rβ) as a standard for IL-3 quantification would result in inaccurate calibration, as the antibodies in IL-3 ELISA kits do not recognize IL-3 Rβ and the molecular weights and epitopes are entirely different.

Best practice:

• For quantification or calibration in ELISA assays targeting mouse IL-3, use purified recombinant mouse IL-3 protein as the standard.
• Recombinant Mouse IL-3 Rβ may be used in receptor binding assays or as a reagent in sandwich ELISAs where the target is IL-3 Rβ, but not for IL-3 quantification.

Summary Table:

If your goal is to quantify mouse IL-3, use recombinant mouse IL-3 protein as your standard. Recombinant Mouse IL-3 Rβ is not suitable for this purpose.

Recombinant Mouse IL-3 Rβ (the beta subunit of the interleukin-3 receptor) has been validated in published research primarily for the following applications:

• Bioassays: Used to assess cell proliferation, particularly in hematopoietic and myeloid cell lines, such as the NFS-60 mouse myelogenous leukemia lymphoblast cell line.
• Cell culture studies: Applied to support the growth, differentiation, and survival of various hematopoietic cell types in vitro.
• Immunoprecipitation and immunoblotting: Utilized in mechanistic studies to investigate receptor interactions, such as the association between IL-3Rβ and c-Kit in EML cells, and to analyze downstream signaling events (e.g., phosphorylation status).
• Transfection experiments: Used in studies where cDNA encoding IL-3Rβ is transfected into cell lines (e.g., 293T cells) to study receptor function and signaling.
• Functional receptor characterization: Employed to distinguish between high-affinity and low-affinity IL-3 receptor complexes and to study their signaling properties in mouse T cell lines.

Supporting details and context:

• Bioassay validation: Numerous studies have used recombinant mouse IL-3 and its receptor components to stimulate cell proliferation and assess cytokine responsiveness in various mouse cell lines, including leukemia and hematopoietic progenitor cells.
• Protein-protein interaction studies: IL-3Rβ has been immunoprecipitated and analyzed by Western blot to study its interaction with other receptors (e.g., c-Kit) and to investigate the molecular mechanisms of cytokine synergy.
• Transfection and expression analysis: Recombinant IL-3Rβ has been expressed in heterologous systems to dissect receptor assembly, ligand binding, and downstream signaling pathways.
• Functional receptor studies: Research has characterized the binding properties and signaling capacity of IL-3Rβ-containing receptor complexes in mouse T cell lines, confirming their role in transmitting growth signals in response to IL-3.

Additional relevant applications:

• Cell differentiation and apoptosis studies: IL-3 and its receptor are involved in regulating cell growth, differentiation, and apoptosis in hematopoietic cells.
• Immunological research: IL-3Rβ is expressed on various immune cell types, including bone marrow progenitors, macrophages, mast cells, eosinophils, megakaryocytes, and basophils, making it relevant for studies on immune cell development and function.

No evidence was found in the provided results for use in diagnostic or therapeutic procedures; all applications are for research purposes only.

To reconstitute and prepare Recombinant Mouse IL-3 Rβ protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Reconstitute the protein in sterile PBS (phosphate-buffered saline) to a concentration of 0.5 mg/mL. If your protocol requires a different concentration, you may adjust within the range of 0.1–1.0 mg/mL as commonly recommended for similar recombinant proteins.
• Gently mix by swirling or gentle pipetting. Do not vortex or mix vigorously, as this can denature the protein.
• Allow the protein to dissolve completely at room temperature. If solubility is an issue, incubate at 4°C overnight.
• For long-term storage, aliquot the reconstituted protein and store at –20°C to –70°C. For short-term use (up to 1 month), store at 2–8°C.
• To prevent protein loss and aggregation, add a carrier protein such as 0.1% BSA or HSA to the solution, especially if storing for extended periods.
• Avoid repeated freeze/thaw cycles by aliquoting into single-use volumes.

Summary protocol:

1. Briefly centrifuge the vial.
2. Add sterile PBS to achieve 0.5 mg/mL (or as required).
3. Gently mix; do not vortex.
4. Allow to dissolve fully.
5. Aliquot and store at –20°C to –70°C (long-term) or 2–8°C (short-term).
6. Add 0.1% BSA/HSA if storing long-term.

Additional notes:

• Always use sterile technique to avoid contamination.
• For cell culture, further dilute the reconstituted stock to the desired working concentration in cell culture medium immediately before use.
• Confirm protein integrity by SDS-PAGE if needed.

These steps ensure optimal solubility, stability, and biological activity of recombinant Mouse IL-3 Rβ for cell culture applications.