Recombinant Human Interleukin-5 (IL5) is expressed in mammalian cells and contains the complete mature protein sequence from amino acids 20 to 134. A 6xHis-tag has been added to the C-terminus to simplify purification and detection processes. SDS-PAGE analysis indicates purity levels above 95%. Endotoxin levels stay below 1.0 EU/µg, which appears to minimize potential experimental interference. The biological activity shows an ED50 of 0.1-0.5 ng/ml when tested in TF-1 cell proliferation assays.
Interleukin-5 (IL5) serves as a cytokine that mainly controls eosinophil regulation and activation, contributing significantly to immune responses. It acts as an essential element in signaling pathways connected to allergic inflammation. Scientists frequently examine IL5 when studying asthma and other conditions involving eosinophils. Research into IL5's mechanisms may offer valuable insights for developing therapeutic approaches that can modify immune responses.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
1. Cell Proliferation and Viability Assays for Eosinophil Research
This recombinant IL-5 protein can stimulate eosinophil proliferation, differentiation, and survival in laboratory settings, since IL-5 is recognized as a primary eosinophil growth factor. The demonstrated biological activity with an ED50 of 0.1-0.5 ng/ml in TF-1 cell proliferation assays shows its functional strength for dose-response experiments. Scientists can apply this protein to explore eosinophil biology, examine how potential inhibitors work, or observe cellular responses under controlled circumstances. High purity levels (>95%) combined with low endotoxin content make it appropriate for delicate cell culture work.
2. IL-5 Receptor Binding and Interaction Studies
The C-terminal 6xHis tag allows for purification and attachment of this IL-5 protein in receptor binding assays and protein-protein interaction experiments. Surface plasmon resonance, bio-layer interferometry, or ELISA-based methods can help researchers characterize IL-5 receptor binding kinetics and affinity. Since the protein comes from a mammalian expression system, it likely maintains proper protein folding and post-translational modifications that are necessary for genuine receptor interactions. Such studies may provide insights into IL-5 signaling mechanisms and support development of receptor antagonists or modulators.
3. Antibody Development and Characterization
This biologically active recombinant IL-5 works well as an antigen for creating and testing anti-IL-5 antibodies in research settings. The high purity and confirmed activity suggest that antibodies developed against this protein should recognize the native, functional form of IL-5. Scientists can incorporate this protein into immunization protocols, ELISA development, Western blot validation, and antibody specificity testing. The His-tag makes protein purification and attachment easier for various antibody screening and characterization procedures.
4. Signal Transduction Pathway Analysis
The confirmed biological activity of this IL-5 protein makes it suitable for examining downstream signaling pathways that activate when IL-5 binds to its receptor. Researchers can apply this protein to stimulate target cells and then analyze phosphorylation events, gene expression changes, and activation of transcription factors like STAT5. The defined ED50 range enables precise dose-dependent studies of signaling cascade activation. These applications might advance understanding of IL-5-mediated cellular responses and help identify potential therapeutic targets within the signaling pathway.
5. Competitive Inhibition and Drug Screening Assays
The established cell proliferation assay system with TF-1 cells creates a foundation for developing competitive inhibition assays to screen potential IL-5 antagonists or modulators. Scientists can use this recombinant protein as a positive control and reference standard in high-throughput screening platforms. The consistent biological activity and low endotoxin levels appear to ensure reliable and reproducible results across multiple experimental conditions. This application supports preclinical drug discovery efforts that target IL-5-mediated pathways.
