Recombinant Human Tumor necrosis factor receptor superfamily member 4 (TNFRSF4) is produced in a mammalian cell expression system, covering amino acids 29-216. This partial protein is C-terminally Fc-tagged and demonstrates a purity greater than 95%, as verified by SDS-PAGE. The endotoxin level is maintained at less than 1.0 EU/µg, assessed by the LAL method. The protein exhibits biological activity, capable of binding Human OX40-Fc with demonstrated affinity constants of 70.3 nM and 165 nM in BLI assays using different biosensors.
TNFRSF4, also known as OX40, belongs to the tumor necrosis factor receptor superfamily and appears to play a critical role in immune system function. It's involved in regulating T-cell activation, promoting cell survival, proliferation, and cytokine production. Given its central role in immune responses, TNFRSF4 has become a significant focus in immunological research, particularly in studies exploring immune modulation and therapeutic interventions.
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. OX40-OX40L Interaction Studies and Binding Kinetics Analysis
This recombinant human OX40-Fc protein can serve as a valuable tool for investigating the molecular interactions between OX40 and its ligand OX40L through surface plasmon resonance, bio-layer interferometry, or other label-free binding assays. The demonstrated binding affinities of 70.3 nM with human OX40L and 165 nM with cynomolgus OX40L provide baseline parameters for comparative studies with mutant variants or under different experimental conditions. Scientists can work with this protein to characterize the thermodynamics and kinetics of OX40-OX40L binding, including association and dissociation rates. The C-terminal Fc tag makes immobilization and detection easier in various binding assays while maintaining the native extracellular domain structure.
2. Competitive Binding Assays for OX40-Targeting Compound Screening
The high-purity OX40-Fc protein with confirmed biological activity can be used in competitive binding assays to screen for small molecules, peptides, or other compounds that modulate OX40-OX40L interactions. Researchers may establish competition assays where test compounds compete with the recombinant protein for binding to immobilized OX40L, which could help identify potential modulators of this signaling pathway. The well-characterized binding affinity provides a reference point for calculating IC50 values and binding competition profiles. The low endotoxin level suggests that observed effects are likely due to specific molecular interactions rather than inflammatory responses.
3. Antibody Development and Characterization Platform
This recombinant OX40 protein works well as an antigen for generating and characterizing anti-OX40 antibodies in research applications. The mammalian expression system appears to ensure proper protein folding and post-translational modifications that closely mimic native human OX40, making it suitable for producing antibodies with physiologically relevant binding properties. Researchers can use this protein in ELISA-based screening of hybridoma clones, antibody affinity measurements, and epitope mapping studies. The Fc tag allows for straightforward purification and immobilization for various immunoassay formats while the confirmed biological activity validates the structural integrity of the antigen.
4. Cross-Species Binding Specificity Studies
The demonstrated differential binding affinities between human OX40L (70.3 nM) and cynomolgus OX40L (165 nM) make this protein valuable for comparative studies of OX40-OX40L interactions across species. Scientists can systematically evaluate binding specificity and affinity variations with OX40L orthologs from different species to understand evolutionary conservation and species-specific differences in this receptor-ligand system. Such studies appear particularly relevant for translational research where animal models are used to study OX40 signaling pathways. The quantitative binding data provides a foundation for predicting cross-reactivity patterns and selecting appropriate animal models for preclinical studies.
5. Protein-Protein Interaction Network Analysis
This biologically active OX40-Fc protein can be used in pull-down assays and co-immunoprecipitation experiments to identify novel binding partners or co-receptors that interact with OX40 in addition to its primary ligand OX40L. Scientists can incubate the protein with cell lysates or purified protein libraries to capture potential interacting molecules, followed by mass spectrometry analysis for identification. The Fc tag makes capture using protein A/G beads straightforward, while the high purity minimizes background binding from contaminants. The confirmed biological activity suggests that identified interactions are likely to be physiologically relevant rather than artifacts of misfolded protein.
