Recombinant Mouse Ig kappa chain V-V region K2 is produced in E. coli, expressing the full length of the mature protein from amino acids 21 to 115. The protein comes with an N-terminal 10xHis-tag and a C-terminal Myc-tag, which appears to make it quite versatile for research work. SDS-PAGE analysis shows purity levels above 85%, suggesting it should work well for most experimental applications that need reliable recombinant protein standards.
The Ig kappa chain V-V region K2 belongs to the immunoglobulin light chain kappa family. These proteins seem to play a crucial role in adaptive immune responses - they're involved in recognizing antigens and form part of the antigen-binding sites on antibodies. Understanding how this protein works may be essential for anyone studying immune system mechanisms or developing therapeutic antibodies.
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.
Mouse Ig kappa chain V-V region K2 is an immunoglobulin variable domain that requires precise folding, proper disulfide bond formation, and specific tertiary structure for its functional activity in antigen binding. The E. coli expression system may not provide the eukaryotic oxidative environment for correct disulfide bond formation, which is critical for immunoglobulin folding. The dual N-terminal 10xHis-tag and C-terminal Myc-tag may cause steric interference with the protein's functional domains, especially given the small size of the variable region (95 aa, ~10.5 kDa). While the full-length mature protein (21-115aa) contains all functional residues, the probability of correct folding with functional antigen-binding activity is low without experimental validation of disulfide bond formation and structural integrity.
1. Immunoglobulin Variable Region Structure-Function Studies
This application carries a significant risk without folding validation. Immunoglobulin variable region function requires precise disulfide bond formation and proper tertiary structure. If correctly folded and active (verified through structural analysis like CD spectroscopy or antigen-binding assays), the protein may be suitable for biophysical studies. If misfolded/inactive (unverified), structural data will yield biologically misleading results due to potential tag interference and improper folding.
2. Antibody Engineering and Development Research
This application requires proper folding validation. Antibody engineering relies on native variable region conformation for meaningful insights. If correctly folded and active (verified), the protein could serve as a reference for scFv or fragment development. If misfolded/inactive (unverified), it may lead to erroneous conclusions in optimization studies.
3. Protein-Protein Interaction Studies
This application carries a high risk without folding validation. Variable region interactions with antigens or other proteins require native conformation. If correctly folded (verified), pull-down assays may identify physiological partners. If misfolded/unverified, there is a high risk of non-specific binding or tag-mediated artefacts.
4. Tag-Based Assay Development and Validation
This application is highly suitable as it focuses on tag detection rather than protein function. The dual tags allow for optimization of ELISA, Western blot, or purification protocols without relying on native protein folding. The known molecular weight and high purity make it an ideal positive control for technical assays.
Final Recommendation & Action Plan
The E. coli-expressed mouse Ig kappa variable region with dual tags may not be properly folded for functional applications due to potential disulfide bond issues and tag interference. Begin with folding validation using techniques like circular dichroism spectroscopy, disulfide bond analysis, and antigen-binding assays before considering Applications 1, 2, and 3. Application 4 (tag-based assay development) can proceed immediately as it is independent of protein function. For reliable variable region research, use mammalian expression systems or implement refolding protocols to ensure native conformation, and consider tag removal for functional studies.
