This recombinant chimeric Dpo4 protein comes from E. coli expression and contains the complete 1-353 amino acid sequence. The N-terminal 6xHis-tag makes purification and detection more straightforward. SDS-PAGE analysis shows the protein achieves over 90% purity, which appears to provide reliable performance for research work. This product is intended strictly for research purposes.
Dpo4 represents a DNA polymerase that plays a key role in translesion DNA synthesis. This process allows DNA replication to proceed past damaged sites that would normally cause the machinery to stall. As a Y-family polymerase, it likely contributes to how cells maintain their genomic integrity. Research into Dpo4's function and molecular interactions may be crucial for understanding DNA repair mechanisms and mutagenesis.
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 Purification and Biochemical Characterization Studies
The N-terminal 6xHis tag allows for straightforward purification through immobilized metal affinity chromatography (IMAC). This makes the recombinant Dpo4 well-suited for detailed biochemical studies. Scientists can examine the protein's biophysical characteristics—thermal stability, oligomerization patterns, and how it binds cofactors. Techniques like differential scanning fluorimetry, analytical ultracentrifugation, and isothermal titration calorimetry become feasible options. The >90% purity should be sufficient for most biochemical assays that need clean protein preparations. This chimeric version could serve as a useful model for studying how engineered polymerase variants differ from their wild-type counterparts.
2. Antibody Development and Immunological Studies
Scientists can use this recombinant chimeric Dpo4 as an immunogen to create polyclonal or monoclonal antibodies targeting this specific engineered construct. The 6xHis tag simplifies immobilization on nickel-coated surfaces, which streamlines antibody screening and ELISA-based studies of binding specificity and affinity. These antibodies might prove valuable for detecting and studying chimeric Dpo4 constructs across different experimental setups. The synthetic nature of this construct appears particularly useful for developing antibodies that can tell engineered variants apart from native polymerase forms.
3. Protein-Protein Interaction Studies
Pull-down assays can take advantage of the His-tagged protein to identify potential binding partners or cofactors that interact with this chimeric Dpo4. The tag enables attachment to nickel-based resins, which can then capture interacting proteins from cell lysates or purified protein mixtures. Surface plasmon resonance (SPR) or bio-layer interferometry experiments may help quantify binding kinetics and affinities with known DNA polymerase-associated factors. Such interaction studies could reveal how the chimeric modifications impact the protein's capacity to form functional complexes.
4. Comparative Structural and Functional Analysis
This chimeric construct offers an interesting opportunity for comparative studies examining how engineered modifications alter protein structure and stability compared to parental Dpo4 variants. Limited proteolysis experiments, hydrogen-deuterium exchange mass spectrometry, or cross-linking studies could map conformational changes that result from the chimeric design. The purified protein works well for thermal shift assays and chemical denaturation studies to measure stability differences. These comparative approaches might advance our understanding of structure-function relationships in engineered DNA polymerases.
