Recombinant Mouse Prohibitin (Phb) is produced in an E.coli expression system and contains a partial sequence spanning amino acids 174 to 272. The protein carries an N-terminal 10xHis-tag, which helps with purification and detection. SDS-PAGE analysis shows the product achieves greater than 85% purity, which appears adequate for most research applications. It's supplied as a research-use-only reagent, though no endotoxin level has been specified.
Prohibitin represents a highly conserved protein that seems central to cellular processes like cell cycle regulation and apoptosis. The protein plays what appears to be a crucial role in maintaining mitochondrial integrity and participates in various signaling pathways. Research often focuses on Prohibitin's potential involvement in cell proliferation and aging, making it an important target for those trying to understand cellular homeostasis.
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 Pull-Down Assays
The N-terminal 10xHis-tagged recombinant mouse Prohibitin fragment (174-272aa) can be immobilized on nickel-affinity resins to hunt down and characterize potential binding partners. This C-terminal region of Prohibitin may retain important protein interaction domains that researchers can probe using cell lysates or purified protein libraries. The His-tag makes purification and immobilization straightforward, which is helpful for both qualitative and quantitative interaction studies. Mass spectrometry analysis of pulled-down complexes might reveal novel interacting proteins specific to this particular Prohibitin domain.
2. Antibody Development and Validation
This recombinant Prohibitin fragment could work as an immunogen for generating domain-specific antibodies against the 174-272aa region of mouse Prohibitin. The purified protein, with its greater than 85% purity, likely provides sufficient quality for immunization protocols in rabbits or other host species. Researchers can then validate the resulting antibodies using the same recombinant protein in ELISA, Western blot, and other immunoassays. Domain-specific antibodies like these would be valuable research tools for studying where Prohibitin localizes and how its expression patterns change.
3. Structural and Biophysical Characterization Studies
The recombinant Prohibitin fragment works well for detailed structural analysis using techniques such as circular dichroism spectroscopy, dynamic light scattering, and nuclear magnetic resonance. These studies would provide insights into how this specific C-terminal domain of mouse Prohibitin folds and maintains stability. The His-tagged protein can be easily purified to the homogeneity that biophysical measurements typically require. Comparing results with full-length Prohibitin or other fragments could reveal structural features unique to specific domains.
4. In Vitro Functional Domain Mapping
This C-terminal fragment (174-272aa) can help map specific functional domains within mouse Prohibitin through comparative biochemical assays. Researchers can test the recombinant protein alongside other Prohibitin fragments or deletion mutants to figure out which activities this specific region retains or loses. Competition assays using this fragment might help identify the minimal binding regions that specific Prohibitin interactions actually need. Studies like these would contribute to understanding how structure and function relate within the Prohibitin protein family.
