Recombinant Human Tumor necrosis factor receptor superfamily member 5 (CD40) is produced in a mammalian expression system, which appears to provide high fidelity to native protein structures. The product consists of the extracellular domain (21-193aa) and is C-terminally Fc-tagged, ensuring stability and ease of detection. With a purity greater than 95% as confirmed by SDS-PAGE, this protein exhibits biological activity, measurable by its binding in a functional ELISA. The endotoxin level is controlled to be less than 1.0 EU/µg, as determined by the LAL method.
CD40 plays a critical role in the immune system. It's primarily known for mediating immune and inflammatory responses. As a member of the tumor necrosis factor receptor superfamily, CD40 is involved in various cellular processes, including B cell activation and regulation of immune responses. Its interactions and signaling pathways make it a significant focus in immunological research, particularly in studies related to immune regulation and potential 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. CD40-CD40L Binding Interaction Studies
This recombinant CD40 extracellular domain can be used to investigate the molecular mechanisms of CD40-CD40L binding interactions in vitro. The demonstrated binding activity with an ED50 of 0.45 μg/ml provides a quantitative basis for dose-response studies and competitive binding assays. Researchers can use this protein to screen potential CD40-CD40L interaction modulators or to characterize binding kinetics using surface plasmon resonance or other biophysical techniques. The high purity and low endotoxin levels make it suitable for sensitive biochemical assays requiring precise measurements.
2. Antibody Development and Characterization
The Fc-tagged CD40 extracellular domain serves as an excellent immunogen and screening target for developing anti-CD40 antibodies. The mammalian expression system likely ensures proper protein folding and post-translational modifications that are crucial for maintaining native epitopes. Researchers can use this protein in ELISA-based screening of hybridoma supernatants or phage display libraries to identify CD40-specific antibodies. The C-terminal Fc tag makes immobilization and detection straightforward in various immunoassay formats.
3. Fc-Tag Mediated Pull-Down Assays
The C-terminal Fc tag enables efficient capture using Protein A or Protein G resins for pull-down experiments to identify CD40-interacting proteins. This approach can be used to study CD40 signaling complexes and discover novel binding partners in cell lysates or purified protein mixtures. The high purity of the recombinant protein minimizes background interactions. Meanwhile, the biological activity suggests that captured proteins represent physiologically relevant interactions. Such studies may provide insights into CD40-mediated signaling pathways in immune cell research.
4. Functional ELISA Development
Based on the validated activity testing method, this CD40 protein can be used to develop standardized functional ELISA assays for research applications. The established binding parameters with CD40L provide a reference point for assay optimization and quality control. Researchers can adapt this system to study CD40L expression levels in cell culture supernatants. They might also evaluate the functional activity of recombinant CD40L preparations. The consistent ED50 value enables reproducible quantitative measurements across different experimental conditions.
5. Structural and Biophysical Characterization Studies
The properly folded extracellular domain expressed in mammalian cells provides what appears to be an ideal substrate for structural biology studies of the CD40 receptor. Researchers can use this protein for crystallization trials, NMR spectroscopy, or cryo-electron microscopy studies to understand CD40 structure-function relationships. The biological activity confirms proper folding, while the high purity ensures minimal interference in biophysical measurements. Such studies may contribute to understanding the molecular basis of CD40 receptor activation and conformational changes upon ligand binding.
