Recombinant Rat Lysosomal acid lipase/cholesteryl ester hydrolase (Lipa) is produced in E. coli and comes with an N-terminal 6xHis-tag. The protein spans the complete mature sequence (26-397 amino acids) and shows a purity level exceeding 85%, as confirmed by SDS-PAGE. This product is strictly for research purposes and cannot be used in diagnostic or therapeutic applications.
Lysosomal acid lipase (Lipa) plays a central role in lipid metabolism by catalyzing the breakdown of cholesteryl esters and triglycerides inside lysosomes. This function appears critical for maintaining proper cellular cholesterol balance, which is why researchers increasingly focus on this enzyme when studying metabolic pathways and related disorders. The way Lipa processes lipids makes it particularly relevant for investigating lipid storage diseases and associated biological processes.
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 rat Lipa protein works well as an immunogen for creating polyclonal or monoclonal antibodies that specifically target rat lysosomal acid lipase. The N-terminal 6xHis tag makes purification straightforward and allows for easy immobilization during antibody screening. Since the protein includes the mature sequence (26-397aa), it represents the biologically active form, which may be ideal for producing antibodies that recognize the native enzyme in rat tissues. These antibodies could prove useful for Western blotting, immunohistochemistry, and immunofluorescence work in rat model systems.
2. Protein-Protein Interaction Studies
The 6xHis tag simplifies pull-down experiments aimed at finding potential binding partners or regulatory proteins that interact with rat Lipa. Researchers can attach the recombinant protein to nickel-affinity resins and expose it to rat tissue lysates or cell extracts to capture any interacting proteins. This strategy might help reveal the molecular mechanisms that control Lipa function and regulation within lysosomal lipid metabolism. Having the full-length mature protein likely preserves all potential interaction sites needed for these binding experiments.
3. Comparative Species Analysis
This rat Lipa protein opens up possibilities for comparative biochemical studies alongside human or mouse Lipa orthologs, potentially revealing species-specific differences in enzyme behavior. Scientists can examine structural variations, stability patterns, and biochemical properties across species using methods like circular dichroism spectroscopy, thermal stability testing, and controlled proteolysis experiments. The recombinant format allows for side-by-side comparisons under identical conditions. Such analyses may provide insights into evolutionary conservation and species-specific modifications of this important lysosomal enzyme.
4. Inhibitor Screening and Binding Studies
The purified recombinant protein functions as a suitable target for screening potential small molecule inhibitors or examining how known Lipa modulators bind. Researchers can measure binding strength using surface plasmon resonance, isothermal titration calorimetry, or fluorescence-based binding methods. The 6xHis tag allows for consistent protein attachment during these experiments, while the high purity (>85%) should provide reliable and repeatable data. These investigations could advance our understanding of Lipa regulation and help identify research tools for controlling its activity in cell-based studies.
