Recombinant Aspergillus restrictus Ribonuclease mitogillin (ret) is produced in E. coli and contains the full-length mature protein from amino acids 28 to 176. The protein includes an N-terminal 10xHis-tag and a C-terminal Myc-tag, which makes purification and detection more straightforward. SDS-PAGE analysis indicates the protein's purity exceeds 85%, suggesting it may be suitable for reliable research applications. This product is intended for research use only.
Ribonuclease mitogillin from Aspergillus restrictus appears to be a ribonuclease enzyme recognized for its role in cleaving RNA molecules. Researchers commonly study it for its enzymatic activity and potential applications in molecular biology research. Understanding how such ribonucleases work and what they do could provide insights into RNA processing and degradation—processes that seem crucial in various cellular and molecular pathways.
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 Dual-Tag Pull-Down Assays
The N-terminal His-tag and C-terminal Myc-tag offer versatile handles for investigating potential binding partners of ribonuclease mitogillin in fungal cellular extracts or purified protein libraries. This dual-tag system allows for sequential purification steps. It may also help confirm interactions through independent capture methods. Researchers can immobilize the protein via either tag to nickel-affinity or anti-Myc antibody matrices and identify co-purifying proteins through mass spectrometry analysis. This approach appears particularly valuable for understanding the cellular context and regulatory networks that might involve this ribonuclease in fungal systems.
2. Antibody Development and Immunoassay Optimization
The recombinant protein with 85% purity serves as what seems to be an appropriate immunogen for generating polyclonal or monoclonal antibodies specific to Aspergillus restrictus ribonuclease mitogillin. The dual-tag configuration could enable comprehensive antibody characterization through tag-specific controls and competition assays. Researchers can develop ELISA-based detection systems using the Myc-tag for capture and test antibodies for detection. This approach likely helps with antibody screening and validation processes. The purified recombinant protein also provides what appears to be a reliable standard for quantitative immunoassays in fungal research applications.
3. Comparative Structural and Biochemical Analysis
The mature protein region (28-176aa) expressed in E. coli provides material for comparative studies with other fungal ribonucleases through techniques such as circular dichroism spectroscopy, dynamic light scattering, and limited proteolysis mapping. Researchers can investigate the protein's stability under various buffer conditions, pH ranges, and temperature profiles to understand its biochemical properties better. The His-tag makes rapid purification possible for multiple analytical preparations, while the defined expression region allows for direct comparison with other characterized ribonuclease domains from related species.
4. Tag-Mediated Protein Localization and Trafficking Studies
The C-terminal Myc-tag may enable researchers to track the recombinant protein's behavior in cell-free systems or permeabilized cell models using anti-Myc antibodies and fluorescent detection methods. This application could prove valuable for studying potential cellular targeting mechanisms or membrane association properties of the ribonuclease when introduced into fungal cell extracts or reconstituted cellular systems. The dual-tag system provides internal controls for specificity. It also allows for orthogonal detection methods in complex experimental setups, though the effectiveness might vary depending on the specific experimental conditions.
