Recombinant Human Stromal cell-derived factor 1 protein (CXCL12) is produced in an E. coli expression system and represents a partial sequence spanning amino acids 22 to 89. This tag-free protein achieves a purity level exceeding 97%, as confirmed by SDS-PAGE analysis. It appears to be fully biologically active, with activity validated via a chemotaxis bioassay using activated human peripheral blood T-lymphocytes, effective at concentrations between 20-80 ng/ml. The endotoxin level remains below 1.0 EU/µg, as determined by the LAL method.
CXCL12, also known as Stromal cell-derived factor 1, plays what seems to be a crucial role in cellular signaling. This chemokine functions primarily in the immune system, directing the movement and localization of cells through chemotaxis. CXCL12 is integral to processes such as hematopoiesis and organ development, which makes it a significant focus in research related to cell migration and immune response pathways.
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. T-lymphocyte Chemotaxis Assays
This recombinant CXCL12 protein can be used to study T-lymphocyte migration patterns in controlled in vitro environments. Given its demonstrated biological activity in chemotaxis bioassays using PHA and rHuIL-2 activated human peripheral blood T-lymphocytes at concentrations of 20-80 ng/ml, researchers may be able to establish standardized migration assays. The high purity (>97%) and low endotoxin levels make it suitable for sensitive cell-based experiments where contamination could confound results. This application would likely prove valuable for investigating immune cell trafficking mechanisms and screening potential modulators of lymphocyte migration.
2. CXCR4 Receptor Binding Studies
The biologically active CXCL12 protein can serve as a ligand in receptor binding assays to characterize CXCR4 interactions. Researchers might use this protein in competitive binding experiments, saturation binding studies, or kinetic analyses to determine binding affinities and receptor occupancy. The defined concentration range of biological activity (20-80 ng/ml) provides a reasonable starting point for dose-response studies. Such experiments would contribute to understanding CXCR4 pharmacology and could potentially support the development of receptor-targeting compounds.
3. Cell Signaling Pathway Analysis
This recombinant protein can be used to investigate downstream signaling cascades triggered by CXCL12-CXCR4 interactions in various cell types. Researchers can treat cells with the biologically active protein and analyze phosphorylation events, second messenger generation, or gene expression changes. The low endotoxin content helps ensure that observed cellular responses are specifically attributable to CXCL12 signaling rather than inflammatory contamination. Time-course and dose-response experiments using the established active concentration range would help elucidate the temporal dynamics of CXCL12-induced signaling.
4. Antibody Development and Validation
The high-purity, tag-free CXCL12 protein represents what appears to be an ideal antigen for generating and characterizing anti-CXCL12 antibodies. The protein can be used for immunization protocols, ELISA development, and antibody specificity testing. Since the protein spans amino acids 22-89 of the mature human CXCL12, it contains key epitopes relevant for antibody recognition. The biological activity confirmation suggests that antibodies developed against this protein are likely to recognize the native, functional form of CXCL12.
5. Protein-Protein Interaction Studies
This biologically active CXCL12 can be used in pull-down assays, surface plasmon resonance, or other biochemical techniques to identify and characterize protein interactions beyond the canonical CXCR4 receptor. The tag-free nature of the protein minimizes potential artifacts from fusion tags that might interfere with native protein interactions. Researchers can investigate binding partners, co-receptors, or regulatory proteins that may modulate CXCL12 function. The established biological activity serves as a quality control measure to help ensure the protein maintains its native conformation during interaction studies.
