Recombinant Rat Galectin-4 (Lgals4) is produced in an E.coli expression system, containing amino acids 1-324 to create the full-length protein. The product comes tag-free, which appears to preserve its native form, and shows a purity level greater than 85% as confirmed by SDS-PAGE analysis. It's designed for research use only, with what seems to be a focus on precision and quality for various experimental applications.
Galectin-4 is a carbohydrate-binding protein that likely plays a significant role in cell-cell and cell-matrix interactions. Research suggests it's involved in several biological processes, including modulation of immune responses and regulation of cell growth and apoptosis. As a member of the galectin family, this protein may be crucial in research aimed at understanding its function in cellular communication and signaling pathways.
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.
Rat Lgals4 is a tandem-repeat galectin containing two carbohydrate recognition domains (CRDs) that requires precise folding, proper disulfide bond formation, and correct domain organization for its carbohydrate-binding activity. The E. coli expression system can produce soluble galectins and supports proper disulfide bond formation, which is favorable for this protein family. The tag-free design eliminates potential steric interference with the CRDs. While the expression system is suitable for galectin production, experimental validation remains essential to confirm structural integrity and carbohydrate-binding functionality.
1. Carbohydrate-Binding Specificity Studies
This application is highly suitable if proper folding is verified. Galectin-4's carbohydrate-binding function requires precise CRD formation. If correctly folded and active (validated through binding assays), the protein is excellent for glycan array and SPR studies. If misfolded/unverified, binding studies will yield biologically meaningless results.
2. Antibody Development and Validation
This application is highly suitable regardless of folding status. Antibody development relies on antigenic sequence recognition rather than functional conformation. The full-length protein provides comprehensive epitope coverage for generating specific antibodies against galectin-4.
3. Protein-Protein Interaction Studies
This application requires proper folding validation. Galectin-4 interactions are often carbohydrate-mediated and require functional CRDs. If correctly folded (verified), the protein may identify physiological interaction partners. If misfolded/unverified, there is a risk of non-specific binding or failure to replicate genuine interactions.
4. Structural and Biophysical Characterization
These studies are essential priority applications for determining folding status. Techniques should include analytical ultracentrifugation to assess dimerization state, circular dichroism spectroscopy to evaluate secondary structure, and thermal shift assays to determine stability. The tag-free design is advantageous for high-resolution structural studies.
5. Cell-Based Functional Assays
This application carries a significant risk without functional validation. Cell-based assays require native protein conformation and carbohydrate-binding capability. If correctly folded and active (verified), limited functional studies may be possible. If misfolded/inactive (unverified), cellular assays will yield biologically misleading results.
Final Recommendation & Action Plan
The E. coli expression system with tag-free design is favorable for producing functional galectin-4, but experimental validation of structural integrity and carbohydrate-binding activity is essential before functional applications. Begin with Application 4 (Structural Characterization) to assess folding quality through analytical ultracentrifugation, CD spectroscopy, and validate carbohydrate-binding capability using standard galectin ligands. Once proper folding and functional activity are verified, proceed cautiously with Applications 1, 3, and 5. Application 2 (antibody development) can proceed immediately. Always include appropriate carbohydrate-binding controls and validate key findings with native protein preparations when possible.
