Recombinant Rat C-X-C motif chemokine 10 protein (Cxcl10) gets expressed in E. coli and covers the complete mature protein sequence, running from amino acids 22-98. The tag-free protein shows high purity—greater than 95% when analyzed through SDS-PAGE. Biological activity appears to remain intact, as confirmed through chemotaxis bioassays using human CXCR3 transfected HEK293 cells at concentrations between 10-50 ng/ml. Endotoxin levels stay controlled below 1.0 EU/µg using the LAL method.
Cxcl10, which people also call Interferon gamma-induced protein 10 (IP-10), acts as a chemokine in immune responses. Its main job seems to be directing immune cell movement, especially by latching onto the CXCR3 receptor. This influences how T cells, NK cells, and other leukocytes migrate and get activated. Because of this, Cxcl10 likely plays an important part in different inflammatory and immune control pathways, which makes it a key focus for immunology researchers.
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. CXCR3 Receptor Binding and Signaling Studies
This recombinant rat CXCL10 protein is confirmed to be biologically active via human CXCR3 (active at 10-50 ng/ml) and suitable for receptor binding studies. However, researchers should validate binding characteristics with rat CXCR3 since the activity was demonstrated using human CXCR3-transfected cells. The high purity (>95%) supports reliable binding assays, but species-specific differences in receptor affinity may exist between rat CXCL10 and human/rat CXCR3 variants.
2. Cell Migration and Chemotaxis Assays
The protein is appropriate for chemotaxis studies, but the activity validation used human CXCR3 in HEK293 cells, not rat immune cells. Researchers should establish dose-response relationships in rat-specific lymphocyte populations (e.g., activated T cells, NK cells) that naturally express CXCR3 to confirm physiological relevance. The concentration range may need adjustment for primary rat cells.
3. Antibody Development and Validation
This high-purity, full-length mature CXCL10 (22-98aa) serves as an excellent antigen for antibody development. However, antibodies should be validated against native rat CXCL10 from inflammatory sources to ensure recognition of physiologically relevant forms, as E. coli expression lacks mammalian post-translational modifications that may affect some epitopes.
4. Protein-Protein Interaction Studies
The biologically active CXCL10 is suitable for interaction studies, but the full functional validation is limited to CXCR3 binding. Researchers exploring interactions beyond CXCR3 should validate that the partial sequence (lacking the signal peptide) maintains all relevant interaction domains present in native, secreted CXCL10.
5. Comparative Species Analysis and Cross-Reactivity Studies
The demonstrated activity on human CXCR3 makes this protein valuable for cross-species studies, but researchers should note that structural differences between rat and human CXCL10 may affect functional comparisons. Parallel assays with human CXCL10 should be conducted using the same experimental system to properly interpret species-specific functional differences.
Final Recommendation & Action Plan
This recombinant rat CXCL10 is a well-characterized reagent suitable for the proposed applications, but requires species-specific validation. First, confirm activity in rat CXCR3-expressing primary cells (T cells, NK cells) to complement the human CXCR3 transfection data. For binding studies, determine species-specific kinetic parameters separately for rat and human CXCR3. When developing antibodies, validate cross-reactivity with native rat CXCL10 from interferon-stimulated tissues. The high purity and low endotoxin support cell-based applications, but researchers should note that the concentration range (10-50 ng/ml) was established in a transfected system and may need optimization for primary cells. For comparative studies, include both rat and human CXCL10 proteins tested in parallel to account for assay variability. The E. coli expression produces a non-glycosylated protein, but the demonstrated bioactivity confirms proper folding for core CXCR3 interactions. Always include appropriate species-matched controls when making functional comparisons.
