This recombinant Mycobacterium tuberculosis Immunogenic protein MPT64 gets expressed in a yeast system and covers the full length of the mature protein from amino acids 24 to 228. An N-terminal 6xHis-tag is included to make purification and detection more straightforward. SDS-PAGE analysis confirms the protein shows purity greater than 90%, which should provide reliable performance in research applications.
MPT64 appears to be a well-characterized immunogenic protein from Mycobacterium tuberculosis that researchers have studied extensively for its role in the immune response to tuberculosis. It represents a significant component in understanding how hosts and pathogens interact and seems crucial for developing diagnostic tools and vaccines. The research community has found that MPT64 helps clarify tuberculosis pathogenesis and immunity, though much remains to be discovered.
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 Characterization
This recombinant MPT64 protein may serve as an immunogen for generating polyclonal or monoclonal antibodies specific to Mycobacterium tuberculosis. The high purity (>90%) and N-terminal 6xHis tag likely make it suitable for immunization protocols and subsequent antibody purification using affinity chromatography. Since the expressed mature protein region (24-228aa) represents the native antigenic form, antibodies generated should recognize the natural MPT64 protein. These antibodies could become valuable research tools for studying M. tuberculosis infection mechanisms and protein localization studies.
2. ELISA-Based Binding Assays
The N-terminal 6xHis tag allows direct immobilization of the protein onto nickel-coated ELISA plates for screening potential binding partners or inhibitors. Researchers might use this system to investigate protein-protein interactions involving MPT64 or to screen compound libraries for molecules that bind to this mycobacterial protein. The high purity level should minimize background interference in binding assays. This approach may provide insights into MPT64's molecular interactions within the mycobacterial proteome or with host cell components, though some interactions could be missed due to the artificial conditions.
3. Immunoblotting and Protein Detection Studies
This recombinant protein works as a positive control and molecular weight standard in Western blot experiments investigating MPT64 expression in M. tuberculosis cultures or clinical isolates. The known protein length and His-tag allow for precise identification using both anti-MPT64 antibodies and anti-His tag antibodies. Researchers can use this protein to validate antibody specificity and optimize detection protocols for studying MPT64 expression under different growth conditions or stress responses.
4. Affinity Purification and Pull-Down Experiments
The 6xHis tag makes it possible to immobilize MPT64 onto nickel-affinity resins for pull-down experiments to identify interacting proteins from M. tuberculosis lysates or host cell extracts. This approach might help clarify the protein interaction network involving MPT64 during mycobacterial infection or metabolism. The high purity of the recombinant protein should reduce non-specific binding, improving the reliability of identifying genuine interaction partners through mass spectrometry analysis of pulled-down complexes. However, some physiologically relevant interactions may be lost when the protein is removed from its native cellular context.
