Recombinant Human Thrombopoietin (THPO) is produced in a mammalian cell expression system, which appears to ensure proper folding and post-translational modifications. This full-length mature protein spans amino acids 22 to 353 and carries a 6xHis tag at both ends. The purity exceeds 95% based on SDS-PAGE verification, and it shows biological activity with an ED50 of 0.55 ng/ml in cell proliferation assays using MO7E cells. Endotoxin levels stay below 1.0 EU/µg, as measured by the LAL method.
Thrombopoietin (THPO) serves as a key growth factor that regulates platelet production. It's central to hematopoiesis—specifically in how megakaryocytes proliferate and differentiate. These are the progenitor cells that generate platelets. Understanding THPO may help researchers grasp disorders affecting platelet counts, like thrombocytopenia and thrombocythemia. This makes it valuable for hematological research.
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. Megakaryocyte Differentiation and Platelet Production Studies
This recombinant human THPO can help investigate how megakaryocytes differentiate and how platelets form in laboratory settings. The demonstrated biological activity with an ED50 of 0.55 ng/ml in MO7E cells gives researchers a solid dose-response reference point. Scientists might use this protein to explore the molecular mechanisms driving thrombopoiesis—things like gene expression changes, signaling pathway activation, and the cellular shape changes that happen as megakaryocytes mature.
2. Cell Proliferation and Viability Assays
The proven activity in MO7E human megakaryocytic leukemic cells makes this protein suitable for standardized proliferation assays and dose-response work. Those dual His-tags allow for straightforward detection and measurement in experimental systems. The high purity (>95%) should deliver consistent results between experiments. This application seems particularly useful for screening studies or when comparing different megakaryocytic cell lines.
3. Protein-Protein Interaction Studies
Both N-terminal and C-terminal His-tags make pull-down assays and affinity purification experiments possible for identifying THPO-binding proteins and receptor interactions. Researchers could use this tagged protein in co-immunoprecipitation studies or surface plasmon resonance experiments to characterize binding kinetics and spot novel interaction partners. The mammalian expression system likely preserves proper protein folding and post-translational modifications that may be crucial for authentic protein interactions.
4. Antibody Development and Validation
This highly pure recombinant THPO works well as an antigen for developing and testing anti-THPO antibodies in research applications. The dual His-tagging makes it easy to attach to various surfaces for ELISA development, Western blot validation, and antibody specificity testing. The confirmed biological activity provides an additional functional readout for checking whether newly developed antibodies interfere with THPO's biological function.
5. Signal Transduction Pathway Analysis
The biologically active THPO can be used to dissect downstream signaling cascades that activate when it binds to receptors in megakaryocytic cells. Researchers might use this protein to study JAK-STAT pathway activation, PI3K/AKT signaling, and other intracellular responses that THPO stimulation triggers. The low endotoxin level (<1.0 EU/μg) helps ensure that observed cellular responses are actually due to THPO activity rather than inflammatory contamination.
