Recombinant Human Antithrombin-III (SERPINC1) is produced through a baculovirus expression system and contains the complete protein sequence spanning amino acids 1 to 464. The protein includes an N-terminal 6xHis tag, which simplifies the purification process. SDS-PAGE analysis confirms that purity levels exceed 85%. This product is designed for research use only and should not be used for diagnostic or therapeutic purposes.
Antithrombin-III appears to be one of the most important serine protease inhibitors in blood coagulation regulation. The protein primarily inhibits thrombin and several other enzymes within the coagulation cascade, which likely helps maintain proper hemostatic balance. Research involving this protein may provide valuable insights into coagulation disorders and anticoagulation strategies, particularly regarding how blood clotting mechanisms work and how they're controlled.
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.
ATIII is a glycoprotein serine protease inhibitor (serpin) that requires complex post-translational modifications, including N-linked glycosylation for correct folding, secretion, and functional interaction with heparin and coagulation factors (e.g., thrombin, factor Xa). The baculovirus expression system supports partial glycosylation and disulfide bond formation, though its glycan patterns differ from those in human cells. Thus, this recombinant ATIII is likely properly folded and capable of partial or full biological activity, depending on how closely insect cell glycosylation mimics human glycoforms.
1. In Vitro Protein-Protein Interaction Studies
This recombinant human ATIIII can be used to study its binding interactions with coagulation factors such as thrombin and factor Xa, and to assess how cofactors like heparin modulate these interactions. If the protein retains proper folding and glycosylation, it can be used for quantitative binding studies (e.g., SPR, BLI) to determine kinetic constants and conformational effects. If bioactivity has not been verified, this protein should be used mainly for qualitative or comparative interaction assays rather than for quantitative affinity measurements. The 6xHis tag facilitates immobilization on nickel-coated surfaces for such binding experiments.
2. Antibody Development and Characterization
The recombinant full-length ATIII is suitable for antibody generation and immunoassay validation, particularly because it contains multiple epitopes distributed throughout the molecule. If folding and glycosylation are native-like, antibodies raised using this protein are likely to recognize both conformational and linear epitopes. If partially misfolded or glycosylation-deficient, it remains suitable for producing linear-epitope antibodies for applications such as Western blotting and ELISA, though such antibodies may not recognize native ATIII in physiological samples.
3. Biochemical Structure-Function Analysis
This recombinant ATIII can be used to investigate structural stability, folding behavior, and conformational transitions typical of serpin proteins. If correctly folded, it is valuable for biophysical studies (e.g., circular dichroism, fluorescence, DLS) examining how heparin binding alters its conformation. If post-translational modifications differ from native ATIII, the protein remains suitable for comparative studies or structure-refinement efforts to evaluate the influence of glycosylation on folding and stability. The His-tag enables efficient purification and immobilization for such studies.
4. Pull-Down Assays for Cofactor Identification
This recombinant ATIII can be used for nickel-affinity pull-down assays to capture potential cofactors or interacting proteins from biological lysates. If the protein maintains correct folding, these experiments may reveal physiologically relevant binding partners such as heparin-binding proteins or serine proteases. If folding or glycosylation is suboptimal, the protein can still serve for in vitro binding screening or identification of potential non-covalent interactors, though results should be interpreted cautiously regarding biological relevance.
5. Comparative Protein Analysis Studies
This recombinant protein provides a useful reference for comparing ATIII variants, including mutant or differently expressed forms. If expressed in mammalian systems, human ATIII typically exhibits full glycosylation and maximal activity; thus, this baculovirus-expressed protein serves as a baseline control for understanding how non-human glycosylation impacts structural and functional properties. It can be applied in side-by-side biochemical and immunological analyses to assess protein expression quality and biological consistency.
Final Recommendation & Action Plan
The recombinant human antithrombin-III (SERPINC1) expressed in the baculovirus system is very likely properly folded and partially to fully active, though its glycosylation profile may differ from the human native form. It is therefore suitable for structural characterization, antibody production, qualitative binding assays, and comparative protein studies. However, for quantitative enzyme inhibition or cofactor-binding kinetics, activity verification should be performed prior to use. If activity is confirmed, prioritize coagulation factor interaction studies and kinetic assays; if not, employ the protein for antibody generation, structural investigations, and quality benchmarking across expression systems.
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