Recombinant Rat Alpha-1-acid glycoprotein (Orm1) is expressed in E. coli and includes an N-terminal 6xHis tag that makes purification more straightforward. The protein covers the complete mature sequence from amino acids 19 to 205. SDS-PAGE analysis shows the product is greater than 85% pure, which appears sufficient for most research applications. This protein is intended for research use only and not for diagnostic or therapeutic purposes.
Alpha-1-acid glycoprotein is an acute-phase plasma protein that seems primarily involved in modulating immune responses. It likely plays an important role in inflammatory processes and is known to bind various ligands, which may affect their distribution and availability. As a major serum protein, Orm1 is frequently studied for its contribution to immune modulation and its potential as a biomarker in research settings.
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. Protein-Protein Interaction Studies Using His-Tag Pull-Down Assays
The N-terminal 6xHis tag on this recombinant rat Orm1 protein allows for nickel-affinity based pull-down experiments to identify potential binding partners. Researchers can immobilize the protein on nickel-coated beads or columns and incubate it with rat tissue lysates or purified proteins to capture interacting molecules. This approach may be particularly valuable for studying Orm1's role in acute phase response pathways and identifying novel regulatory proteins. While the >85% purity level appears sufficient for these interaction studies, contaminating proteins can typically be distinguished through mass spectrometry analysis of pulled-down complexes.
2. Antibody Development and Validation
This recombinant rat Orm1 protein serves as a useful antigen for generating specific antibodies against rat alpha-1-acid glycoprotein. The purified protein can be used for immunizing animals to produce polyclonal antibodies or screening hybridomas for monoclonal antibody production. The His-tagged protein also functions as a positive control in ELISA-based antibody characterization assays, allowing researchers to determine antibody specificity, affinity, and cross-reactivity. Since the mature protein sequence (19-205aa) represents the native form, it should be suitable for developing antibodies that recognize endogenous rat Orm1.
3. Biochemical Characterization and Ligand Binding Studies
Researchers can use the recombinant protein in biochemical assays to characterize the binding properties of rat Orm1 with various ligands, including drugs, lipids, and other small molecules. Fluorescence polarization, surface plasmon resonance, or isothermal titration calorimetry experiments can determine binding kinetics and thermodynamic parameters. The protein's purity level of >85% is likely adequate for these quantitative binding studies, though appropriate controls should be included to account for any remaining contaminants.
4. Comparative Species Analysis in Acute Phase Response Research
This rat Orm1 protein can be used alongside human and mouse orthologs in comparative studies to understand species-specific differences in acute phase protein function. Scientists can perform side-by-side binding assays, stability studies, or structural comparisons to identify conserved and divergent functional domains. The recombinant protein may enable controlled experiments where species-specific effects can be isolated from other variables, potentially contributing to translational research that bridges rodent models and human physiology.
