Recombinant Human Probable E3 ubiquitin-protein ligase HERC1 is produced in an E.coli expression system. The protein features a 10xHis-tag at the N-terminus and a Myc tag at the C-terminus. This partial protein covers amino acids 3975-4360 and reaches a purity level greater than 85%, as confirmed by SDS-PAGE analysis. It's designed for research purposes and appears to offer a solid tool for studying protein interactions and modifications, with low endotoxin levels that may be beneficial for cell-based studies.
HERC1 belongs to the ubiquitin-protein ligase family. It seems to play an important role in the ubiquitination process, which targets proteins for degradation through the proteasome pathway. This protein likely participates in various cellular pathways and processes, which makes it particularly interesting for research into protein regulation and cellular homeostasis. Studying HERC1 could potentially improve our understanding of cellular mechanisms and how proteins are managed throughout their lifecycle.
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 Validation
This recombinant HERC1 fragment (amino acids 3975-4360) may work well as an immunogen for creating specific antibodies against HERC1's C-terminal region. The dual His and Myc tags make purification and detection more straightforward during antibody screening. Scientists can potentially use this protein fragment to test antibody specificity through Western blot, ELISA, and immunoprecipitation assays. The high purity level (greater than 85%) suggests reliable antigen presentation for both polyclonal and monoclonal antibody production, though results may vary depending on the immunization protocol.
2. Protein-Protein Interaction Studies
The His and Myc tags should allow for pull-down assays to identify potential binding partners that interact with this particular C-terminal domain of HERC1. Scientists can immobilize the tagged protein on suitable matrices and then screen cell lysates or protein libraries for interacting molecules. This strategy appears especially useful for mapping the interaction network of HERC1's C-terminal region, which might contain important regulatory or substrate recognition domains—though the functional significance of identified interactions would need further validation.
3. Structural and Biochemical Characterization
This defined protein fragment offers a manageable substrate for biophysical studies. These might include circular dichroism spectroscopy, dynamic light scattering, and analytical ultracentrifugation to examine its folding properties and oligomerization state. The tags help with protein detection and quantification during purification and analysis steps. Scientists can explore the stability, solubility, and conformational properties of this HERC1 domain under different buffer conditions and temperatures, though the behavior of this fragment may not fully represent the full-length protein.
4. ELISA-Based Quantitative Assays
The dual-tagged HERC1 fragment could serve as a standard or control protein in sandwich ELISA assays for detecting and measuring HERC1 levels in biological samples. The His tag permits oriented immobilization on nickel-coated plates, while the Myc tag provides an additional detection epitope. This application appears useful for researchers studying HERC1 expression levels across different cell types, developmental stages, or disease conditions in preclinical research settings, although antibody cross-reactivity and sample matrix effects would need careful consideration.
