Recombinant Human Ras-related protein Rab-10 (RAB10) is produced using a baculovirus expression system and spans the complete protein sequence (1-200 amino acids). The protein comes with an N-terminal 10xHis-tag and a C-terminal Myc-tag, which makes purification and detection more straightforward. SDS-PAGE analysis shows purity levels above 85%, making this product suitable for research applications.
Rab-10 belongs to the Rab family of small GTPases, proteins that appear to be crucial regulators of how membranes move around inside cells. It seems particularly important in endocytic recycling, where it influences how proteins get transported and sorted within cells. This involvement in cellular transport pathways has made Rab-10 an interesting target for researchers studying these mechanisms.
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.
Based on the provided information, the recombinant Human RAB10 is expressed in a Baculovirus system (insect cells), which is a eukaryotic expression system capable of supporting proper folding and post-translational modifications for eukaryotic proteins like RAB10. The protein is full-length (1-200aa), which includes all functional domains, and the dual tags (N-terminal 10xHis and C-terminal Myc) are designed to minimize interference. However, since activity is unknown, the protein cannot be assumed to be correctly folded or bioactive without functional validation (e.g., GTP binding or effector interaction assays). While the Baculovirus system increases the probability of correct folding, experimental confirmation is essential to ensure bioactivity.
1. Protein-Protein Interaction Studies Using Pull-Down Assays
The dual tags enable technical feasibility for pull-down assays, but if RAB10 is misfolded, it may not interact physiologically with binding partners (e.g., effectors or regulators), leading to non-specific or false interactions. The tags facilitate purification and detection, but results should be interpreted cautiously until folding is validated. If correctly folded, this application is valuable; otherwise, it may yield misleading data.
2. Antibody Development and Validation
This application is well-supported. The recombinant RAB10 can serve as an immunogen for antibody generation, as antibodies may recognize linear epitopes even if the protein is misfolded. The high purity (85%) and tags aid in purification and screening. However, if misfolded, antibodies may not recognize conformational epitopes of native RAB10, so validation against endogenous protein is recommended.
3. Biochemical Characterization and Stability Studies
This application is appropriate and should be prioritized to assess folding. Techniques like thermal stability assays or circular dichroism can directly evaluate protein conformation. The tags facilitate detection and purification, and these studies are valuable even if the protein is inactive, as they characterize the recombinant product.
4. ELISA-Based Quantitative Assays
The dual-tag system allows for sandwich ELISA development, but if RAB10 is misfolded, conformational epitopes may be altered, leading to inaccurate quantification. The assay may work for detection, but results should be validated with native RAB10 or activity checks to ensure reliability. It should be noted that folding validation is needed for accurate quantitative use.
Final Recommendation & Action Plan
Given the uncertainty in folding and bioactivity, it is recommended to first perform biochemical and functional validation to confirm the protein's conformation and activity. Use techniques such as size-exclusion chromatography (for oligomeric state), circular dichroism (for secondary structure), and GTP-binding assays to assess folding and function. If validated, the protein can be reliably used for all described applications; if not, focus on antibody development and biochemical characterization, while avoiding interaction studies that require native conformation. Always include appropriate controls, such as native RAB10 or activity standards, in experiments. The Baculovirus expression system is favorable, but activity confirmation remains crucial for functional applications.
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