Recombinant Mouse Cystatin-C (Cst3) is produced in E. coli and includes an N-terminal 10xHis-tag along with a C-terminal Myc-tag, which makes purification and detection more straightforward. The protein represents the complete mature form, covering amino acids 21-140. SDS-PAGE analysis confirms purity levels above 85%, making this product well-suited for research applications that demand highly specific and consistent recombinant protein.
Cystatin-C appears to be a cysteine protease inhibitor that researchers study extensively for its regulatory function in proteolytic pathways. It seems crucial for maintaining protease balance and may have important implications in research focusing on tissue remodeling, inflammation, and cellular homeostasis. Understanding how it interacts with other molecules is likely essential for revealing the mechanisms behind protease-related processes in biological systems.
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. Cysteine Protease Inhibition Assays
This recombinant mouse Cystatin-C works well as a positive control or standard inhibitor in biochemical assays that examine cysteine protease activity. The dual His and Myc tags make purification and detection simpler in enzyme kinetics studies that look at inhibitory effects on cathepsins and other cysteine proteases. Research teams can use this protein to create dose-response curves and calculate IC50 values for protease inhibition experiments. The high purity level (>85%) helps ensure reliable and reproducible results in quantitative enzyme assays.
2. Antibody Development and Validation
The dual-tagged recombinant protein appears to work as an excellent immunogen and positive control for creating mouse Cystatin-C specific antibodies. The N-terminal His tag makes protein purification easier for immunization protocols. Meanwhile, the C-terminal Myc tag offers an additional epitope for validation experiments. Researchers can apply this protein in ELISA development, Western blot optimization, and establishing immunohistochemistry protocols. The mature protein sequence (21-140aa) may represent the biologically relevant form for generating antibodies with physiological specificity.
3. Protein-Protein Interaction Studies
The His-tagged recombinant Cystatin-C can be attached to nickel-affinity matrices for pull-down assays that identify novel binding partners or confirm known interactions. This protein works as bait in biochemical binding studies with tissue lysates or purified proteins to map interaction networks. Co-immunoprecipitation experiments can take advantage of the Myc tag for detection and validation of binding specificity. These interaction studies might provide insights into Cystatin-C's role in cellular processes beyond simple protease inhibition.
4. Structural and Biophysical Characterization
This purified recombinant protein supplies material for structural biology studies, including X-ray crystallography, NMR spectroscopy, and cryo-electron microscopy experiments. The high purity level makes it appropriate for biophysical analyses like dynamic light scattering, circular dichroism spectroscopy, and thermal stability assays. Scientists can examine the protein's folding properties, stability under different conditions, and conformational changes when binding occurs. The dual tags can be used for oriented attachment in surface plasmon resonance studies.
5. Comparative Species Studies
The mouse-specific recombinant Cystatin-C allows comparative biochemical studies between mouse and human versions to understand species-specific differences in protease inhibition profiles. Cross-reactivity studies can be conducted using this protein with proteases from various species to map evolutionary conservation of inhibitory mechanisms. The protein works as a reference standard for developing mouse-specific assays in preclinical research models. Research teams can use this protein to validate how relevant mouse models are for human Cystatin-C related studies.
