Recombinant Human Ephrin-A4 (EFNA4) is produced using a mammalian cell expression system and covers amino acid region 26-171. The partial protein carries a C-terminal 6xHis-Fc tag, which makes purification and detection more straightforward. SDS-PAGE analysis confirms purity levels exceeding 95%. The protein shows biological activity - functional ELISA testing reveals a binding ED50 of 1.5190 ug/ml to Human EphA7-His. Endotoxin levels stay below 1.0 EU/μg.
Ephrin-A4 appears to play an important role in cell signaling, especially during development. As part of the ephrin family, it works with Eph receptors to enable cell-to-cell communication. This interaction seems critical for several pathways that affect how cells stick together, move around, and organize into tissues. These functions make Ephrin-A4 an interesting target for scientists studying how cells develop and communicate.
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. Eph-Ephrin Signaling Pathway Studies
This recombinant EFNA4 protein may prove useful for investigating how Eph-ephrin signaling works at the molecular level in cell culture systems. The binding activity to EphA7 with a defined ED50 value makes it well-suited for dose-response experiments that examine receptor activation and the signaling events that follow. Scientists can study bidirectional signaling - both forward signaling through Eph receptors and reverse signaling through ephrin ligands. High purity and low endotoxin levels should give reliable results in sensitive cell-based experiments.
2. Receptor-Ligand Interaction Analysis
The C-terminal His-Fc tag allows for purification and detection in biochemical assays designed to characterize how Eph and ephrin proteins bind to each other. Surface plasmon resonance (SPR) or bio-layer interferometry (BLI) experiments can use this protein to measure binding kinetics, affinity constants, and how specifically it interacts with different Eph receptor family members. The Fc tag also makes it easy to attach the protein to protein A/G surfaces for binding studies. These interaction studies might provide useful quantitative data about receptor selectivity and how different proteins compete for binding sites within the Eph-ephrin system.
3. Cell Adhesion and Migration Assays
Scientists can attach EFNA4 to culture surfaces to study how it affects cell adhesion, spreading, and migration in laboratory conditions. The protein's biological activity and ability to engage Eph receptors suggests it could be valuable for investigating how cell-to-cell contact influences cellular behavior. Researchers might coat plates or beads with this protein to examine how EFNA4-Eph interactions change cell attachment, detachment, or directional movement. The mammalian expression system likely ensures proper protein folding and modifications that could be important for these cellular responses.
4. Antibody Development and Validation
The recombinant EFNA4 protein works well as an antigen for creating and testing antibodies that specifically recognize human ephrin-A4. High purity and the defined expression region (26-171aa) offer a consistent target for immunization protocols and antibody screening. The His-Fc tag makes purification and immobilization easier for ELISA-based antibody testing, while the demonstrated biological activity suggests the protein maintains proper shape. This protein can help validate antibody specificity, identify binding sites, and create standard curves for quantitative immunoassays.
5. Functional ELISA Development
Building on the activity testing approach described, this protein may be useful for developing and optimizing functional ELISA assays to study Eph-ephrin interactions. The established binding protocol with EphA7 and known ED50 value provide a starting point for creating standardized binding assays. Researchers could potentially use this protein to develop competitive binding assays, screen for small molecule inhibitors, or measure Eph receptor expression levels in cell lysates or tissue samples. The Fc tag helps with detection and the low endotoxin content should support assay reliability.
