Recombinant Human Cartilage intermediate layer protein 1 (CILP) is produced through a baculovirus expression system, covering amino acids 725-1184. The protein comes with an N-terminal 10xHis tag and C-terminal Myc tag for easier detection and purification. This partial protein shows greater than 85% purity when analyzed by SDS-PAGE, which appears to make it appropriate for research applications requiring high-quality protein samples.
Cartilage intermediate layer protein 1 (CILP) is mainly known for its role in cartilage's extracellular matrix. The protein seems to be involved in how cartilage develops and maintains itself, likely playing an important part in keeping the tissue structurally sound and functional. Because CILP participates in cartilage biology, it has become a key target for researchers trying to understand cartilage-related processes and diseases.
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 CILP fragment (amino acids 725-1184) with its dual His and Myc tags may work well as an immunogen for creating specific antibodies against human CILP. The partial protein sequence covers a fairly large chunk of the full-length protein, suggesting it could produce antibodies that recognize native CILP in human cartilage samples. Having both tags makes purification and detection simpler during antibody screening. Researchers might find this protein useful for validating antibody specificity through Western blotting, ELISA, and immunoprecipitation experiments.
2. Protein-Protein Interaction Studies
Both the N-terminal His tag and C-terminal Myc tag allow for pull-down assays to hunt for potential CILP binding partners in cartilage tissue extracts or cell lysates. Since the baculovirus expression system typically produces properly folded proteins with correct post-translational modifications, there's a better chance of keeping native protein interactions intact. Researchers can attach this tagged protein to suitable matrices and screen for interacting proteins using mass spectrometry or Western blot analysis. Such studies could reveal more about how CILP functions in cartilage matrix organization and cellular signaling pathways.
3. ELISA-Based Quantification Assays
The dual-tagged recombinant protein appears suitable as a standard or control in sandwich ELISA assays for measuring CILP levels in biological samples. Having both His and Myc tags gives researchers multiple options for capture and detection antibody strategies, which makes assay design more flexible. This protein could help establish standard curves for quantifying endogenous CILP in synovial fluid, cartilage extracts, or cell culture supernatants. The >85% purity level should be sufficient for creating reliable and reproducible quantification assays in cartilage research.
4. Biochemical Characterization and Stability Studies
This recombinant CILP fragment may prove useful for investigating the protein's biochemical properties - things like thermal stability, pH sensitivity, and how easily it breaks down when exposed to proteases. The dual tagging system makes protein detection and quantification straightforward throughout various experimental conditions using tag-specific antibodies. Researchers can test how different buffer conditions, salt concentrations, or chemical treatments affect protein stability and shape. These kinds of studies would likely add to our understanding of how CILP behaves in the cartilage extracellular matrix environment and help guide future experimental approaches.
