Recombinant Human Hyaluronan and Proteoglycan Link Protein 4 (HAPLN4) is produced in E. coli and spans the complete mature protein sequence from amino acids 30 to 402. The protein carries an N-terminal 10xHis-tag for easier purification and detection. SDS-PAGE analysis shows purity levels above 85%, which appears to provide reliable performance for research work.
HAPLN4 likely plays a crucial role in stabilizing how hyaluronic acid and proteoglycans interact within the extracellular matrix. It seems particularly important for maintaining the structural integrity of cartilage and neural tissues. Research into HAPLN4's molecular pathways may prove essential for understanding how tissues develop and repair themselves.
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 HAPLN4 protein works well as an immunogen for creating both polyclonal and monoclonal antibodies that target human HAPLN4. The N-terminal 10xHis tag makes purification straightforward and allows for easy immobilization during antibody screening. Researchers can also use it as a positive control in Western blots, ELISAs, and immunofluorescence experiments to confirm antibody specificity. The 85%+ purity level should be sufficient for most immunization protocols and follow-up antibody characterization work.
2. Protein-Protein Interaction Studies
His-tagged HAPLN4 works particularly well in pull-down experiments designed to find new binding partners or verify known interactions with extracellular matrix components. The 10xHis tag allows researchers to attach the protein to nickel-based resins, making it possible to capture interacting proteins from cell lysates or purified protein solutions. This method may help clarify HAPLN4's actual role in organizing the extracellular matrix and reveal how it interacts within proteoglycan complexes. The recombinant protein can also serve as bait in co-immunoprecipitation studies when paired with suitable antibodies.
3. ELISA-Based Quantitative Assays
The purified HAPLN4 protein serves as either a standard or coating antigen in enzyme-linked immunosorbent assays for research purposes. The His tag enables consistent, oriented attachment to nickel-coated plates, which might improve both assay reliability and sensitivity. Researchers can use this protein to build sandwich ELISAs for measuring HAPLN4 concentrations in cell culture media or tissue samples. The known concentration and purity make it well-suited for creating standard curves in quantitative immunoassays.
4. Cell Culture Studies and Extracellular Matrix Research
Researchers can add this recombinant HAPLN4 directly to cell culture systems to examine how it affects cell behavior, adhesion, or movement patterns in vitro. The protein proves useful for studying extracellular matrix assembly by observing cellular responses to added HAPLN4 under controlled culture conditions. Scientists might explore its potential in tissue engineering applications or investigate its broader functions in extracellular matrix biology. Using recombinant protein creates more controlled experimental conditions compared to proteins extracted from tissues, which often show batch-to-batch variation.
