Recombinant Human Interleukin-3 (IL3) is produced in E. coli and represents the full length of the mature protein, spanning amino acids 20-152. It features an N-terminal 6xHis-tag for easier purification. The product achieves high purity of over 95% as verified by SDS-PAGE and maintains an endotoxin level of less than 1.0 EU/µg, making it suitable for research applications. The protein shows biological activity and can bind Human IL-3RA-Fc with affinity constants of 3.89 µM and 3.74 µM in BLI assays.
Interleukin-3 (IL3) is a cytokine that appears to play a significant role in hematopoiesis, particularly in the differentiation and proliferation of hematopoietic stem cells. It participates in signaling pathways that may regulate immune cell development and function. IL3's activity seems critical for maintaining the balance of various blood cell types, which is why it has garnered considerable interest in research focused on immunology and hematological studies.
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. IL-3 Receptor Binding Studies and Affinity Characterization
This recombinant IL-3 protein can be used to investigate the binding kinetics and thermodynamics of IL-3 interactions with its receptor subunits, particularly IL-3RA. The demonstrated binding affinity of approximately 3.8-3.9 μM to IL-3RA-Fc provides a baseline for comparative studies with receptor variants or mutants. Researchers might use surface plasmon resonance, biolayer interferometry, or other biophysical techniques to characterize binding parameters under different conditions. The N-terminal His-tag simplifies protein immobilization and purification for these binding assays.
2. Hematopoietic Cell Culture and Proliferation Assays
Given IL-3's established role as a hematopoietic growth factor, this biologically active recombinant protein could support in vitro studies of hematopoietic progenitor cell proliferation and differentiation. The protein may be useful in colony-forming unit assays, bone marrow culture systems, or studies with hematopoietic cell lines to investigate cellular responses to IL-3 stimulation. The low endotoxin level (less than 1.0 EU/μg) makes it suitable for sensitive cell culture applications where bacterial contamination might confound results.
3. Antibody Development and Immunoassay Standardization
High purity (>95%) and the His-tagged format make this protein an excellent antigen for generating anti-IL-3 antibodies or as a standard in immunoassays. Researchers can use it to develop ELISA protocols, validate antibody specificity, or create calibration curves for IL-3 quantification assays. The His-tag allows easy purification and immobilization on nickel-based surfaces for antibody screening applications.
4. Protein-Protein Interaction Studies
Beyond receptor binding, this IL-3 protein could be used to investigate interactions with other signaling molecules, co-receptors, or regulatory proteins in the IL-3 signaling pathway. Pull-down assays using the His-tag may identify novel binding partners from cell lysates or protein libraries. The confirmed biological activity suggests that the protein maintains its native-like conformation necessary for physiologically relevant protein interactions.
5. Structure-Function Relationship Analysis
This recombinant IL-3 can serve as a reference protein for mutagenesis studies aimed at understanding critical residues for receptor binding and biological activity. Researchers might compare the binding affinity and cellular responses of IL-3 variants to this wild-type protein using the established BLI assay format with IL-3RA-Fc. The mature protein sequence (aa 20-152) represents the functional domain, which appears to make it ideal for systematic structure-activity relationship investigations.
