Recombinant Mouse Stromal cell-derived factor 1 protein (Cxcl12) is produced in E. coli and spans amino acid region 22-89, creating a partial protein without any tags. The protein achieves high purity, with levels exceeding 97% as determined by SDS-PAGE. Testing confirms it remains fully biologically active and shows chemotactic activity in bioassays with human peripheral blood monocytes at concentrations ranging from 50-100 ng/ml. Endotoxin levels stay below 1.0 EU/µg, ensuring minimal contamination.
Stromal cell-derived factor 1 (Cxcl12) functions as a chemokine central to cellular signaling. It plays a key role in chemotaxis, guiding the migration of various cell types throughout the body. This protein participates in multiple biological processes, including hematopoiesis and immune response regulation. Cxcl12 has become widely studied for its involvement in developmental and physiological pathways, making it an important focus for research into cellular communication and migration.
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. Chemotaxis Assay Development and Optimization
This recombinant mouse CXCL12 protein appears well-suited as a positive control and standard reference in chemotaxis assays for studying cell migration responses. Since it demonstrates biological activity in human peripheral blood monocyte chemotaxis at 50-100 ng/ml concentrations, it may provide a reliable tool for establishing dose-response curves and validating experimental conditions. The high purity (>97%) and low endotoxin levels suggest it's appropriate for sensitive cell-based assays where contamination could skew migration results. Researchers might compare chemotactic responses across different cell types and experimental conditions using this protein.
2. CXCR4 Receptor Binding Studies
The biologically active CXCL12 protein can be applied in receptor binding assays to investigate CXCR4 receptor interactions and binding kinetics. Competition binding experiments could be performed using this protein as a competitor against labeled ligands to determine binding affinities and receptor occupancy. The defined expression region (22-89aa) represents the mature, secreted form of the protein, which likely makes it suitable for studying physiologically relevant receptor-ligand interactions. These studies may support research into CXCL12-CXCR4 signaling pathways and receptor pharmacology.
3. Cell Culture Supplementation for Migration Studies
This recombinant protein could be added to cell culture media to create chemokine gradients for studying directional cell migration in vitro. The confirmed biological activity and established concentration range (50-100 ng/ml) offer guidance for creating effective gradients in transwell migration assays or microfluidic devices. The E. coli expression system and tag-free nature mean the protein lacks mammalian post-translational modifications that might interfere with controlled experimental conditions. Researchers can explore how different cell types respond to CXCL12 gradients under various experimental setups.
4. Antibody Development and Validation
The high-purity, tag-free recombinant mouse CXCL12 protein may serve as an effective antigen for developing and characterizing anti-CXCL12 antibodies. The defined amino acid sequence (22-89aa) allows for precise epitope mapping studies and specificity testing of newly developed antibodies. The biological activity confirmation suggests that antibodies developed against this protein target the functionally relevant form of CXCL12. This protein could prove useful in ELISA development, Western blot validation, and immunoassay standardization for CXCL12 detection and quantification.
5. Protein-Protein Interaction Studies
This biologically active CXCL12 protein can be applied in biochemical assays to study its interactions with binding partners beyond CXCR4, such as glycosaminoglycans or other potential regulatory proteins. The tag-free nature eliminates potential interference from fusion tags in binding studies, while the high purity appears to ensure reliable results in pull-down assays or surface plasmon resonance experiments. The low endotoxin content makes it suitable for sensitive biochemical assays where bacterial contaminants might affect protein interactions. These studies could help clarify the molecular mechanisms underlying CXCL12 function and regulation.
