Recombinant Human Kit ligand (KITLG) is produced in an E. coli expression system, covering the amino acid region 26-189. This product is tag-free and shows a purity level exceeding 95%, as verified by SDS-PAGE analysis. It demonstrates biological activity, with an effective dose (ED50) of 1-5 ng/ml, determined through a cell proliferation assay using TF-1 human erythroleukemic cells. The endotoxin level remains below 1.0 EU/µg, as measured by the LAL method.
Kit ligand (KITLG), also known as stem cell factor (SCF), appears to play a crucial role in hematopoiesis. It promotes the survival, proliferation, and differentiation of hematopoietic stem cells. The protein is involved in various signaling pathways and seems essential for the development of melanocytes, germ cells, and interstitial cells of Cajal. Researchers commonly use KITLG to study stem cell biology and related cellular processes.
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. Cell Proliferation and Viability Assays
This recombinant KITLG can stimulate cell proliferation in hematopoietic cell lines and primary cells expressing the KIT receptor. The established ED50 of 1-5 ng/ml in TF-1 cells provides a validated concentration range for dose-response studies. Scientists may use this protein to investigate cell cycle progression, survival signaling pathways, and growth factor dependencies in various blood cell lineages. High purity and low endotoxin levels make it suitable for sensitive cell culture applications where contamination could potentially skew results.
2. Hematopoietic Stem Cell Research
KITLG appears essential for hematopoietic stem cell maintenance and differentiation. This makes the recombinant protein valuable for in vitro stem cell studies. Scientists can use it to maintain primitive hematopoietic progenitors in culture or to study the molecular mechanisms that may govern stem cell fate decisions. The protein can be incorporated into defined media formulations for ex vivo expansion of hematopoietic stem cells from bone marrow or cord blood samples. Its biological activity likely ensures proper receptor activation necessary for studying stem cell biology and hematopoiesis.
3. Signal Transduction Pathway Analysis
This biologically active KITLG serves as a tool to investigate KIT receptor-mediated signaling cascades in various cell types. Scientists can study downstream phosphorylation events, protein-protein interactions, and gene expression changes following receptor activation. The defined concentration range allows for controlled stimulation experiments to map temporal signaling dynamics. Time-course studies using this protein may help clarify the kinetics of pathway activation and identify key regulatory nodes in KIT signaling networks.
4. Antibody Development and Validation
The high-purity, tag-free recombinant KITLG works as an antigen for developing and characterizing anti-KITLG antibodies. Researchers can use it in immunization protocols for monoclonal antibody generation or as a standard in antibody specificity testing. The protein serves as a positive control in immunoassays such as ELISA, Western blotting, and immunoprecipitation experiments. Its biological activity can also validate the functional impact of neutralizing antibodies in cell-based assays.
5. Protein-Protein Interaction Studies
This recombinant KITLG works well in biochemical assays to study its interactions with the KIT receptor and other binding partners. Surface plasmon resonance, bio-layer interferometry, or other biophysical techniques can determine binding kinetics and affinity constants. Scientists may also use the protein in pull-down assays or co-immunoprecipitation experiments to identify novel interacting proteins. Its soluble nature and high purity make it well-suited for structural biology applications and protein complex characterization studies.
