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.
Rat Lysosomal acid lipase/cholesteryl ester hydrolase (Lipa) is a complex eukaryotic enzyme that requires precise folding, proper glycosylation, disulfide bond formation, and lysosomal targeting for its functional activity in lipid metabolism. The E. coli expression system cannot provide the necessary eukaryotic post-translational modifications (particularly glycosylation) and molecular chaperones required for this lysosomal enzyme. While the protein may be soluble, it is highly unlikely to achieve the correct folding, glycosylation pattern, and enzymatic activity needed for functional lipase/cholesteryl ester hydrolase activity.
1. Antibody Development and Validation
Antibody development relies on antigenic sequence recognition rather than functional folding or glycosylation. This recombinant Lipa serves as an excellent immunogen for generating antibodies against linear epitopes of rat lysosomal acid lipase. The full-length mature sequence ensures comprehensive epitope coverage. However, antibodies may not efficiently recognize glycosylation-dependent or conformational epitopes on the native, properly modified enzyme found in physiological conditions.
2. Biophysical Characterization Studies
Basic biophysical characterization can be performed to assess the protein's physical properties. Techniques like circular dichroism spectroscopy can analyze secondary structure content, while size-exclusion chromatography can determine oligomeric state and aggregation behavior. However, these studies will characterize a non-glycosylated, misfolded protein rather than the native enzyme.
Final Recommendation & Action Plan
The E. coli expression system is fundamentally unsuitable for producing a functional version of this complex lysosomal enzyme due to its glycosylation requirements. This recombinant Lipa is primarily suitable for antibody development against linear epitopes (Application 1) and basic biophysical characterization (Application 2), but the characterization data will reflect a non-native protein state. For reliable Lipa functional studies, use mammalian expression systems that support proper glycosylation and lysosomal targeting, or work with native enzyme purified from biological sources where correct post-translational modifications are preserved. Begin with basic biophysical characterization to confirm protein integrity, then proceed with linear epitope antibody production. All functional applications require alternative protein sources with proper glycosylation and folding validation.
