< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
QC176, Quantikine Immunoassay Control Set 643 Human Thrombopoietin - Please Inquire
The Quantikine Human Tpo Immunoassay is a 4.5 hour solid phase ELISA designed to measure human Tpo in cell culture supernates, serum, and plasma. It contains recombinant human Tpo and antibodies raised against the recombinant factor. Results obtained using natural human Tpo showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural Tpo.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
Serum, EDTA Plasma, Heparin Plasma
The recovery of Tpo spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
Platelet-poor EDTA Plasma (n=4)
Platelet-poor Heparin Plasma (n=4)
To assess the linearity of the assay, samples spiked with high concentrations of Tpo were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Thrombopoietin (Tpo), is a key regulator of megakaryocytopoiesis and thrombopoiesis. It is principally produced in the liver and is bound and internalized by the receptor Tpo R/c-mpl. Defects in the Tpo-Tpo R signaling pathway are associated with a variety of platelet disorders. The 353 amino acid (aa) human Tpo precursor is cleaved to yield the 332 aa mature protein. Mature human Tpo shares approximately 70% aa sequence homology with mouse and rat Tpo. It is an 80 - 85 kDa protein that consists of an N-terminal domain with homology to Erythropoietin (Epo) and a C-terminal domain that contains multiple N-linked and O-linked glycosylation sites. Tissue specific alternate splicing of human Tpo generates multiple isoforms with internal deletions, insertions, and/or C-terminal substitutions. Tpo promotes the differentiation, proliferation, and maturation of MK and their progenitors. Several other cytokines can promote these functions as well but only in cooperation with Tpo. Notably, IL-3 independently induces MK development, although its effects are restricted to early in the MK lineage. Tpo additionally promotes platelet production, aggregation, ECM adhesion, and activation. It is cleaved by platelet-derived thrombin following Arg191 within the C-terminal domain and subsequently at other sites upon extended digestion. Full length Tpo and shorter forms circulate in the plasma. The C-terminal domain is not required for binding to Tpo R or inducing MK growth and differentiation. Aside from its hematopoietic effects, Tpo is expressed in the brain where it promotes the apoptosis of hypoxia-sensitized neurons and inhibits neuronal differentiation by blocking NGF induced signaling.
Thrombopoietin receptor (Tpo R), also known as myeloproliferative leukemia protein (c-Mpl), is a type I transmembrane protein that is a member of the hematopoietin/cytokine receptor superfamily. As a consequence of alternative splicing, there are four identified mRNA variants. The functional receptor is encoded by the P isoform of mRNA. The human and mouse receptors share approximately 81% amino acid sequence identity. Tpo R is expressed at low levels in various cell types, including hematopoietic progenitor cells, megakaryocytes and platelets.
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