Recombinant Human Aminopeptidase N (ANPEP) is produced in E. coli with an expression region spanning amino acids 34 to 219. This partial protein carries an N-terminal 10xHis-GST tag and a C-terminal Myc tag, which help with purification and detection. The purity exceeds 85%, as verified by SDS-PAGE analysis, ensuring reliable performance in research applications.
Aminopeptidase N (ANPEP) is an enzyme that appears to play a significant role in peptide substrate metabolism. It cleaves amino acids from the N-terminus of peptide chains—a process that seems important for protein maturation and recycling. ANPEP also participates in various cellular pathways, making it a valuable research target for studying enzymatic function and regulation.
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. Antibody Development and Validation Studies
This recombinant ANPEP fragment (34-219aa) can work as an immunogen or antigen for developing antibodies against human aminopeptidase N. The dual-tagged protein offers multiple detection and purification strategies during antibody screening. Researchers may use this protein in ELISA-based assays to validate antibody specificity and determine binding affinities. The 85% purity level is likely sufficient for immunization protocols and subsequent antibody characterization experiments.
2. Protein-Protein Interaction Studies
The His-GST tag system allows for efficient pull-down assays to identify potential binding partners of the ANPEP extracellular domain. Researchers can immobilize the GST tag on glutathione-sepharose beads to capture interacting proteins from cell lysates or protein libraries. The Myc tag provides an additional detection method for confirming protein presence and integrity during interaction studies. This partial ANPEP construct represents the extracellular region that would be accessible for physiological protein interactions.
3. Structural and Biochemical Characterization
The recombinant protein can be used for basic biochemical analyses. These might include size exclusion chromatography, dynamic light scattering, and thermal stability studies to understand the folding and stability properties of this ANPEP domain. The dual tagging system helps with protein purification and detection during various analytical procedures. Researchers can investigate the biophysical properties of this specific ANPEP region (amino acids 34-219) and compare them with other aminopeptidase family members.
4. Tag-Based Detection System Development
The combination of His, GST, and Myc tags makes this protein valuable for developing and optimizing multi-tag detection systems and purification protocols. Researchers can use this protein as a positive control in Western blotting, immunoprecipitation, and affinity purification experiments. The protein serves as a useful standard for testing the specificity and sensitivity of anti-His, anti-GST, and anti-Myc antibodies across different experimental conditions.
