Recombinant Mouse Fructose-bisphosphate aldolase C (Aldoc) is expressed in yeast and contains the complete mature protein sequence, spanning amino acids 2-363. The protein includes a C-terminal 6xHis tag that simplifies purification and detection processes. SDS-PAGE analysis confirms the purity exceeds 90%, which appears to provide reliable results for research applications. The recombinant protein comes with low endotoxin levels, making it appropriate for sensitive experimental work.
Fructose-bisphosphate aldolase C serves as a key enzyme in the glycolytic pathway. It catalyzes the reversible conversion of fructose-1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The enzyme is mainly found in the central nervous system, where it likely plays an important role in energy metabolism within neuronal cells. This makes it particularly interesting for studies focused on metabolic processes and neurological research.
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. Biochemical Characterization of Aldolase C Enzymatic Properties
This recombinant mouse Aldoc protein offers researchers a way to examine the basic enzymatic properties of fructose-bisphosphate aldolase C. Studies might include kinetic parameters, substrate specificity, and determining optimal reaction conditions. The full-length mature protein from yeast expression appears suitable for measuring aldolase activity using standard spectrophotometric assays that track fructose-1,6-bisphosphate cleavage. The C-terminal 6xHis tag makes protein purification straightforward and allows for immobilization during detailed kinetic studies. Comparing results with other aldolase isoforms may help reveal what makes the C isoform biochemically unique.
2. Antibody Development and Validation Studies
The purified recombinant Aldoc protein works well as an immunogen for creating specific antibodies against mouse aldolase C. Its high purity (>90%) and full-length structure should ensure proper epitope presentation for both polyclonal and monoclonal antibody production. Researchers can use the C-terminal His tag in ELISA-based screening assays to identify and characterize antibody specificity and binding affinity. These antibodies would become valuable research tools for immunoblotting, immunoprecipitation, and immunofluorescence studies that investigate Aldoc expression and where it localizes in cells.
3. Protein-Protein Interaction Studies
The His-tagged recombinant Aldoc can be used in pull-down assays to identify novel protein interaction partners or confirm known interactions with other glycolytic enzymes or regulatory proteins. The tag allows for efficient immobilization on nickel-affinity matrices, which can capture potential binding partners from cell lysates or purified protein preparations. Mass spectrometry analysis of these pulled-down complexes might reveal new insights into aldolase C's role in metabolic enzyme complexes or cellular signaling pathways. This approach seems particularly valuable for studying tissue-specific interactions, given that aldolase C has a distinct expression pattern compared to other aldolase isoforms.
4. Structural and Biophysical Analysis
This full-length recombinant protein provides material for structural biology studies, including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy to understand the three-dimensional architecture of mouse aldolase C. Researchers can use the protein in biophysical characterization experiments such as dynamic light scattering, analytical ultracentrifugation, or thermal stability assays to determine oligomerization states and stability parameters. Comparing structures with aldolase A and B isoforms could reveal the molecular basis for functional differences. The yeast expression system typically produces properly folded eukaryotic proteins that appear suitable for high-resolution structural analysis.
5. Metabolic Pathway Reconstitution Assays
The recombinant Aldoc protein can be incorporated into in vitro glycolytic pathway reconstitution experiments to study metabolic flux and enzyme coordination in controlled biochemical systems. These cell-free systems allow precise manipulation of enzyme concentrations and cofactor availability to understand how aldolase C contributes to overall glycolytic efficiency. The purified protein enables quantitative analysis of metabolic intermediates and can be combined with other recombinant glycolytic enzymes to model tissue-specific metabolic characteristics. Such studies may prove valuable for understanding metabolic regulation and the specific role of aldolase C in different physiological contexts.
