Recombinant Human Galectin-3-binding Protein (LGALS3BP) is produced in E. coli and includes the full-length mature protein sequence spanning amino acids 19 to 585. The protein comes with an N-terminal 6xHis tag, which makes purification and detection more straightforward. SDS-PAGE analysis confirms the product reaches greater than 85% purity - a level that appears adequate for most experimental work. This recombinant protein is intended strictly for research purposes and offers reliable specifications for scientific studies.
Galectin-3-binding protein participates in several cellular processes, particularly cell adhesion and immune response regulation. As a secreted glycoprotein, it interacts with galectins and seems to play a meaningful role in how cells communicate with the extracellular matrix. Scientists are investigating LGALS3BP to better grasp how it contributes to cellular signaling networks, which may help uncover new insights about disease development and potential therapeutic approaches.
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.
Based on the provided information, the recombinant Human LGALS3BP is expressed in E. coli, a prokaryotic system that is generally unsuitable for producing properly folded eukaryotic glycoproteins like LGALS3BP. This protein requires complex glycosylation and proper disulfide bond formation for its native conformation and bioactivity, neither of which E. coli can provide. The protein is expressed as the full-length mature protein (19-585aa) with an N-terminal 6xHis tag, and purity is >85% by SDS-PAGE. However, since activity is unverified and the expression system cannot support proper glycosylation (critical for LGALS3BP's lectin-binding functions), the protein is highly likely to be misfolded, unglycosylated, and inactive. Experimental validation is essential to confirm both proper folding and bioactivity.
1. Protein-Protein Interaction Studies Using His-Tag Pull-Down Assays
The N-terminal 6xHis tag enables technical feasibility for pull-down assays to identify potential binding partners. However, if LGALS3BP is misfolded and unglycosylated (as is likely in E. coli), it will not interact physiologically with true binding partners like galectin-3 or other lectins. Identified interactions may be non-specific artifacts. This application should not be pursued without prior validation of proper folding and glycosylation-mimetic functionality.
2. Antibody Development and Validation
This application is appropriate. The recombinant LGALS3BP can serve as an immunogen for generating antibodies that recognize linear epitopes, even if the protein is misfolded. The His-tag facilitates purification and screening. However, antibodies may not recognize conformational or glycosylation-dependent epitopes of native LGALS3BP in physiological contexts. Validation against glycosylated, native LGALS3BP from mammalian sources is essential.
3. Biochemical Characterization and Stability Studies
This application is well-suited and should be prioritized. Techniques like circular dichroism spectroscopy, size-exclusion chromatography, and thermal stability assays can directly assess the protein's folding state, oligomerization, and stability. These studies are valuable even if the protein is inactive, as they characterize the recombinant LGALS3BP protein itself and can inform about its suitability for other applications.
4. ELISA-Based Quantitative Assays
This application is problematic without proper folding validation. If LGALS3BP is misfolded and unglycosylated, it will not function effectively as a capture agent or standard in ELISA assays designed to detect native, glycosylated LGALS3BP. Antibodies may not recognize the recombinant protein similarly to the native form, leading to inaccurate quantification. This application requires confirmation that the recombinant LGALS3BP protein's immunoreactivity matches native LGALS3BP.
Final Recommendation & Action Plan
Given the high probability of misfolding due to a lack of glycosylation in E. coli, we recommend first performing a comprehensive biophysical characterization (circular dichroism for secondary structure, size-exclusion chromatography for oligomeric state) to assess folding quality. Antibody development can proceed, but resulting antibodies must be validated against native, glycosylated LGALS3BP. Avoid interaction studies and quantitative assays until proper folding is confirmed. For reliable functional studies of LGALS3BP's galectin-binding properties, obtain the protein from a mammalian expression system capable of proper glycosylation. If using this E. coli-expressed protein, limit applications to antibody production (with validation) and biochemical characterization.
