Recombinant Human Intestinal-type alkaline phosphatase (ALPI) is produced in an E. coli expression system, covering amino acids 26 to 500. The protein carries an N-terminal 6xHis-tag for straightforward purification. It arrives with purity levels above 90%, as verified by SDS-PAGE analysis, which appears to support consistent performance in research applications. This product is designed strictly for research use and meets strict quality standards for endotoxin levels.
Intestinal-type alkaline phosphatase (ALPI) is an enzyme that's essential for dephosphorylation processes. It plays an important role in the digestive system by breaking down dietary phosphates. The enzyme participates in various physiological pathways, including nutrient absorption and gut barrier function. ALPI's activity seems vital for maintaining intestinal health and has become a subject of extensive research due to its regulatory roles in the human body.
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. Protein-Protein Interaction Studies Using His-Tag Affinity Purification
The N-terminal 6xHis tag allows for nickel-affinity chromatography-based pull-down experiments to identify potential binding partners of human intestinal-type alkaline phosphatase. Researchers can immobilize the recombinant ALPI on nickel-coated beads or columns and incubate it with cell lysates or purified protein libraries to capture interacting proteins. Bound proteins can then be eluted and analyzed by mass spectrometry to map the ALPI interactome. This approach may be particularly valuable for understanding the regulatory networks and cellular pathways involving ALPI in intestinal physiology.
2. Antibody Development and Validation
The >90% pure recombinant ALPI protein appears to serve as an excellent immunogen for generating polyclonal or monoclonal antibodies specific to human intestinal-type alkaline phosphatase. The protein can be used to immunize laboratory animals or in phage display systems for antibody selection. Additionally, the purified protein provides what seems to be a reliable positive control for validating antibody specificity through Western blotting, ELISA, and immunoprecipitation assays. The His-tag makes purification and immobilization easier for antibody screening applications.
3. Biochemical Characterization and Enzyme Kinetics Analysis
Despite the absence of confirmed biological activity testing, this recombinant ALPI can be subjected to comprehensive biochemical analysis to determine its enzymatic properties. Standard alkaline phosphatase substrates such as p-nitrophenyl phosphate can be used to assess catalytic activity under various pH and temperature conditions. Researchers can systematically evaluate the protein's stability, optimal reaction conditions, and kinetic parameters. These studies would likely provide fundamental biochemical data for understanding the enzymatic properties of the intestinal-type alkaline phosphatase isoform.
4. Comparative Structural and Functional Studies
The recombinant ALPI protein makes comparative analysis with other alkaline phosphatase isoforms possible to identify structural and functional differences specific to the intestinal type. The protein can be used in limited proteolysis experiments, chemical cross-linking studies, and hydrogen-deuterium exchange mass spectrometry to probe its structural features. Comparative enzyme assays using different substrates and inhibitors may reveal functional distinctions between ALPI and other alkaline phosphatase family members, contributing to understanding isoform-specific roles.
