Recombinant Human Protein disulfide-isomerase (P4HB) is produced in E. coli and provided as a full-length mature protein spanning amino acids 18-508. The product includes an N-terminal 6xHis-tag for straightforward purification and detection. SDS-PAGE analysis confirms a purity level exceeding 90%, meeting high-quality standards for research applications. Endotoxin levels are kept minimal to support various experimental protocols.
Protein disulfide-isomerase (P4HB) appears to play a central role in disulfide bond formation, which seems essential for proper protein folding. The enzyme catalyzes thiol-disulfide exchange reactions, contributing to protein structural stability. P4HB represents a key component in cellular processes related to protein synthesis and maintenance, making it an important target for studies examining protein folding and cellular homeostasis.
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 Folding and Disulfide Bond Formation Studies
This recombinant P4HB can investigate protein disulfide isomerase activity in vitro using model substrate proteins containing disulfide bonds. Scientists may examine the enzyme's role in promoting proper protein folding by monitoring disulfide bond formation and rearrangement in target proteins. The His-tagged format allows for straightforward purification and immobilization on nickel-based resins for controlled enzymatic assays. Such applications could prove valuable for understanding cellular protein quality control mechanisms and oxidative protein folding pathways.
2. Biochemical Characterization and Enzyme Kinetics Analysis
The purified recombinant protein enables detailed biochemical studies to determine kinetic parameters, optimal reaction conditions, and cofactor requirements for P4HB enzymatic activity. Scientists can perform substrate specificity assays using various disulfide-containing peptides and proteins to map the enzyme's catalytic preferences. High purity levels make this protein suitable for accurate quantitative measurements and reproducible kinetic studies across different experimental conditions.
3. Antibody Development and Immunoassay Applications
This recombinant P4HB may serve as an immunogen for generating specific antibodies against human protein disulfide isomerase in research applications. The His-tag streamlines purification for antibody production protocols and enables development of sandwich ELISA assays for detecting P4HB in biological samples. The purified protein can also function as a positive control and standard in Western blotting, immunoprecipitation, and other antibody-based detection methods.
4. Protein-Protein Interaction Studies
His-tagged recombinant P4HB can be used in pull-down assays to identify and characterize protein binding partners and substrate interactions. Researchers might immobilize the protein on nickel affinity matrices to capture interacting proteins from cell lysates or purified protein mixtures. This approach appears useful for mapping the P4HB interactome and understanding its role in protein folding networks within the endoplasmic reticulum environment.
5. Structural Biology and Biophysical Analysis
High-purity recombinant protein is suitable for structural studies including X-ray crystallography, NMR spectroscopy, and cryo-electron microscopy to elucidate P4HB's three-dimensional structure. Biophysical characterization techniques such as dynamic light scattering, circular dichroism spectroscopy, and thermal stability assays may provide insights into protein conformation and stability. The His-tag can be used for oriented immobilization in surface plasmon resonance studies to analyze binding kinetics with various substrates and inhibitors.
