Recombinant Human Repulsive guidance molecule A (RGMA) is produced in E. coli and includes the full-length mature protein spanning amino acids 169 to 424. The protein features an N-terminal 10xHis-tag and a C-terminal Myc-tag, which help with purification and detection processes. SDS-PAGE analysis confirms a purity level exceeding 85%, making it appropriate for various research applications.
RGMA belongs to the repulsive guidance molecule family. It appears to play a critical role in axon guidance and neural development. The protein is involved in signaling pathways that may influence neural patterning and regeneration, which has made it a subject of considerable interest in neurobiology research. Understanding RGMA's interactions and functions could provide insights into nervous system development and potential therapeutic approaches for neural repair.
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 Studies
This recombinant RGMA protein can work as an immunogen or antigen for developing monoclonal or polyclonal antibodies against human RGMA. The dual tagging system - with its N-terminal His-tag and C-terminal Myc-tag - offers multiple epitopes for antibody validation through Western blot, ELISA, and immunoprecipitation assays. The 85% purity level appears sufficient for immunization protocols and subsequent antibody characterization studies. Researchers can likely use this protein to generate research-grade antibodies for studying RGMA expression and localization across different cell types and tissues.
2. Protein-Protein Interaction Studies
The His-tagged RGMA protein can be immobilized on nickel-affinity matrices for pull-down assays to identify potential binding partners from cell lysates or purified protein libraries. Meanwhile, the Myc-tag allows detection and confirmation of successful protein capture in these interaction studies. This approach may help researchers investigate RGMA's molecular interactions and identify novel binding partners that could be involved in guidance signaling pathways. The expressed region (169-424aa) represents the mature protein form, which suggests it's suitable for physiologically relevant interaction studies.
3. ELISA-Based Binding Assays
Both tags work together to support the development of sandwich or direct ELISA formats for quantitative binding studies. The protein can be coated onto ELISA plates via the His-tag while the Myc-tag serves as a detection epitope using anti-Myc antibodies. This dual-tag system provides flexibility in assay design. It enables researchers to study RGMA binding to various ligands, receptors, or other molecules of interest. The 85% purity is likely adequate for establishing reliable and reproducible ELISA-based assays.
4. Biochemical Characterization and Stability Studies
This recombinant RGMA protein can be used for comprehensive biochemical analysis. This includes molecular weight confirmation, thermal stability assessment, and buffer optimization studies. The defined expression region and dual tagging allow for precise protein identification and tracking during various experimental conditions. Researchers can investigate protein stability under different pH, salt, and temperature conditions to establish optimal storage and handling protocols. These studies may provide fundamental biochemical data necessary for downstream applications and assay development.
