Recombinant Mouse Chymotrypsin-like elastase family member 3B (Cela3b) is produced using an E.coli expression system, covering the full length of the mature protein from amino acids 28 to 269. The protein carries an N-terminal 10xHis-tag and a C-terminal Myc-tag, which should make detection and purification more straightforward. SDS-PAGE analysis shows the product achieves greater than 90% purity, suggesting it's well-suited for various research applications.
Cela3b appears to be a serine protease that's involved in protein digestion and processing. It likely plays a critical role in breaking down dietary proteins within the digestive system. The protein belongs to the trypsin family of proteases, which are known for their ability to cleave peptide bonds. Scientists frequently study this protein when investigating digestive enzyme function and regulation, making it an important target in enzymology and digestive system research.
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 Studies
This recombinant mouse Cela3b protein may work well as an immunogen or antigen for developing specific antibodies against chymotrypsin-like elastase family member 3B. The dual-tagged design (N-terminal His-tag and C-terminal Myc-tag) should allow for straightforward purification and detection during immunization protocols and subsequent antibody validation assays. With >90% purity, it appears suitable for generating high-quality polyclonal or monoclonal antibodies. Scientists can then apply this protein in ELISA, Western blot, and immunoprecipitation experiments to assess antibody specificity and binding affinity.
2. Protein-Protein Interaction Studies
The dual-tag system (His and Myc tags) seems ideal for pull-down assays and co-immunoprecipitation experiments aimed at identifying potential binding partners of Cela3b. Scientists can immobilize the protein through the His-tag on nickel-based resins. Alternatively, they might use anti-Myc antibodies for immunoprecipitation studies with mouse tissue lysates or cell extracts. Since the mature protein region (28-269aa) represents the biologically relevant form, it should be appropriate for studying physiological protein interactions. These experiments could help reveal the molecular pathways and regulatory networks that involve Cela3b.
3. Biochemical Characterization and Enzyme Kinetics Analysis
While biological activity hasn't been tested, this recombinant protein may prove useful for establishing and optimizing enzymatic assays for chymotrypsin-like elastase activity using synthetic substrates. The high purity (>90%) and full-length mature protein structure suggest it's suitable for detailed biochemical characterization studies. Scientists can investigate optimal buffer conditions, pH requirements, and cofactor dependencies for potential enzymatic activity. The protein might also serve as a standard or control when comparing it with other elastase family members.
4. Structural and Biophysical Studies
The high purity level of this recombinant Cela3b protein suggests it could work well for various biophysical characterization techniques. These might include dynamic light scattering, circular dichroism spectroscopy, and thermal stability analysis. Such studies could provide insights into protein folding, secondary structure content, and stability under different conditions. The dual-tag system allows for easy purification and concentration of the protein for biophysical experiments. This characterization data would likely prove valuable for understanding the structural properties of this elastase family member and optimizing storage and handling conditions.
