Recombinant Human TIMD4 is produced using a mammalian expression system and spans amino acids 25 to 314 of the complete protein. The product includes a C-terminal 6xHis-hFc1 tag that aids in purification and detection. SDS-PAGE analysis confirms the protein achieves high quality with purity levels exceeding 85%. This product is designed for research use only and appears to deliver consistent results across different experimental applications.
TIMD4 - short for T-cell immunoglobulin and mucin domain-containing protein 4 - plays what seems to be a critical role in immune regulation. The protein is involved in clearing apoptotic cells, which is essential for maintaining immune homeostasis. Researchers studying immune response pathways and cell clearance mechanisms may find this protein particularly useful for gaining deeper insights into these fundamental biological processes.
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.
The recombinant human TIMD4 (25–314aa) expressed in a mammalian cell system and tagged with C-terminal 6xHis-hFc1 is highly likely to be correctly folded and biologically active in terms of ligand recognition. This fragment encompasses the entire extracellular region of TIMD4, which includes the IgV-like domain and part of the mucin-like region—the key regions responsible for phosphatidylserine (PS) binding and cell-surface interactions. The mammalian expression system ensures native disulfide bond formation, glycosylation, and overall proper tertiary structure, all of which are essential for maintaining the biological activity of TIMD4. Therefore, this recombinant protein may retain native-like conformation and functional binding properties, though specific activity (e.g., PS binding) must still be experimentally validated before claiming full biological equivalence to membrane-bound TIMD4.
1. Protein-Protein Interaction Studies
This recombinant TIMD4 fragment (25–314aa) can be used to study ligand binding interactions, especially with phosphatidylserine and potential protein partners involved in apoptotic cell recognition. If the protein retains its native folding—as expected from mammalian expression—it can be used in pull-down assays, surface plasmon resonance (SPR), or biolayer interferometry (BLI) to examine binding affinity and kinetics. If folding or glycosylation is altered during production, it may still serve as a screening reagent for identifying potential linear or glycan-independent interactions, though results may not fully reflect physiological binding. Overall, this construct is suitable for ligand-binding and receptor-mapping assays, provided its conformation is verified.
2. Antibody Development and Characterization
This recombinant protein is well-suited for generating and screening antibodies specific to the extracellular domain of TIMD4. If properly folded, it will display native conformational epitopes, allowing for the production of antibodies that recognize TIMD4 in cell-based or tissue-based contexts. If misfolded or partially denatured, antibodies generated will mainly target linear epitopes, making them more useful for Western blot or ELISA detection. The C-terminal His-hFc1 tag simplifies both antigen purification and antibody validation, providing a versatile platform for antibody development workflows.
3. Structural and Biophysical Analysis
This TIMD4 fragment may serve as material for structural and biophysical characterization. If properly folded (as expected from mammalian expression), it can be subjected to X-ray crystallography, cryo-EM, or NMR to determine the architecture of the extracellular domain and its interaction surfaces. If folding or glycosylation is not fully native, the protein remains valuable for CD spectroscopy, DLS, or DSC to assess secondary structure content and stability. The Fc fusion promotes dimerization and stability, aiding in crystallization or higher-order assembly studies, but should be considered when interpreting oligomeric states.
4. Cell-Based Binding and Uptake Assays
The mammalian-expressed TIMD4 protein is suitable for cell-based assays examining binding to PS-expressing cells or potential receptor interactions. If correctly folded and glycosylated, it can function as a biologically relevant probe in flow cytometry, confocal microscopy, or receptor competition assays using the Fc tag for detection. If folding or glycosylation deviates from the native form, binding results should be interpreted cautiously, as structural or glycan differences may alter receptor affinity. Overall, the recombinant Fc fusion protein is highly appropriate for cell binding and uptake studies, particularly with validation of activity via PS-coated surfaces or apoptotic cell binding controls.
Final Recommendation & Action Plan
The recombinant human TIMD4 expressed in mammalian cells and fused with a C-terminal 6xHis-hFc1 tag is very likely correctly folded and functionally competent for ligand-binding applications. It represents the entire extracellular domain, making it a biologically relevant reagent for interaction, antibody, and cell-binding studies. However, functional validation of PS-binding activity or structural integrity (e.g., via CD spectroscopy or binding assays) is recommended before using it in quantitative or mechanistic analyses.
