Recombinant Human MGAT4A is expressed in E.coli and covers amino acid region 93-535, representing a partial length version of the protein. The product includes an N-terminal 6xHis tag for easier purification and detection. SDS-PAGE analysis confirms the protein reaches purity levels above 85%. This preparation is designed for research use only and appears to deliver consistent performance across different experimental setups.
MGAT4A represents a key glycosyltransferase that modifies N-glycans and plays an important role in creating complex-type oligosaccharides. The enzyme's activity affects N-glycan branching patterns, which may influence protein folding and cell-cell interactions. Studying MGAT4A could help researchers better understand glycosylation pathways and how they relate to cellular function and disease 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.
1. Biochemical Characterization and Enzyme Kinetics Studies
Researchers can use this recombinant MGAT4A protein to explore the enzyme's basic biochemical properties - things like optimal pH, temperature stability, and what cofactors it needs to function. The N-terminal 6xHis tag makes purification straightforward and allows for protein immobilization during kinetic experiments. Scientists can measure Km and Vmax values across different substrate concentrations to get a sense of the enzyme's catalytic efficiency. The partial protein construct (93-535aa) likely retains the catalytic domain, which should make it useful for core enzymology work.
2. Antibody Development and Validation
This purified recombinant protein works well as an immunogen for creating polyclonal or monoclonal antibodies against human MGAT4A. The >85% purity appears sufficient for standard immunization protocols and antibody testing. The 6xHis tag simplifies purification and can be helpful in tag-based ELISA assays when screening and validating antibody specificity. Such antibodies would likely prove valuable for Western blotting, immunoprecipitation, and tracking protein location within cells.
3. Protein-Protein Interaction Studies
Scientists can immobilize the 6xHis-tagged MGAT4A on nickel-affinity matrices for pull-down assays aimed at finding potential binding partners or regulatory proteins. The recombinant protein may serve as bait in biochemical screens using cell lysates or purified protein collections. Co-immunoprecipitation experiments that take advantage of the His tag could help reveal protein complexes that involve MGAT4A in glycosylation networks. Even this partial construct might preserve important protein interaction domains needed for studying how the enzyme is regulated.
4. Substrate Specificity and Inhibitor Screening
This recombinant enzyme allows researchers to systematically test substrate preferences and identify specific glycoprotein targets for N-acetylglucosaminyltransferase activity. Having purified protein enables controlled in vitro assays for testing various potential substrates and optimizing reaction conditions. Scientists can use this protein to screen for potential inhibitors or compounds that modulate MGAT4A activity in drug discovery efforts. The standardized protein preparation should provide reproducible results when comparing different experimental conditions.
