Recombinant Rat MICOS complex subunit Mic60 (Immt) is produced through an in vitro E.coli expression system and contains amino acids 34 to 609, which creates a partial protein product. The protein includes an N-terminal 10xHis-tag that helps with purification and detection. SDS-PAGE analysis shows the protein achieves a purity level exceeding 85%, making it what appears to be a solid choice for research applications targeting mitochondrial studies.
The MICOS complex subunit Mic60—also known as mitochondrial contact site and cristae organizing system—likely plays a crucial role in maintaining mitochondrial architecture. This protein seems particularly important in forming cristae junctions and organizing the mitochondrial inner membrane overall. Mic60's involvement in these processes may highlight why it's become significant in studies examining mitochondrial function and dynamics.
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 for MICOS Complex Assembly
This recombinant Mic60 fragment can be applied in pull-down assays to identify and characterize interactions with other MICOS complex subunits or mitochondrial proteins. The N-terminal His-tag allows for immobilization on nickel-affinity matrices during co-immunoprecipitation experiments. Scientists can examine the binding domains and stoichiometry of MICOS complex formation with this purified protein component. The partial length construct (34-609aa) might reveal specific interaction domains within the Mic60 protein sequence.
2. Antibody Development and Validation
The purified recombinant protein works well as an antigen for generating specific antibodies against rat Mic60. Researchers can use the His-tagged protein in ELISA-based screening of hybridoma clones or polyclonal sera to identify high-affinity, specific antibodies. Scientists may validate antibody specificity through Western blot analysis with this recombinant protein as a positive control. The defined protein region (34-609aa) allows for mapping epitope specificity in antibody characterization studies.
3. Structural and Biophysical Characterization
This recombinant Mic60 fragment can support structural studies including circular dichroism spectroscopy, dynamic light scattering, and analytical ultracentrifugation to characterize protein folding and oligomerization states. The purified protein enables investigation of thermal stability and conformational changes under various buffer conditions. Scientists can perform limited proteolysis experiments to identify structured domains within the 34-609aa region. The His-tag helps with protein purification for high-concentration samples that biophysical analyses typically require.
4. In Vitro Functional Assays for Mitochondrial Membrane Organization
The recombinant protein can be integrated into liposome-based assays to study its role in membrane organization and cristae junction formation. Scientists can investigate the protein's ability to interact with cardiolipin or other mitochondrial lipids through lipid-binding assays. The purified Mic60 fragment enables reconstitution experiments to study its contribution to mitochondrial membrane architecture in defined in vitro systems. These studies may provide insights into the molecular mechanisms behind cristae junction stability and organization.
