Quantikine Immunoassay Control Set 934 Human Thrombopoietin
Quantikine Immunoassay Control Set 934 Human Thrombopoietin Summary
INTENDED USE & DESCRIPTION
For use as quantitative controls for the determination of cytokine concentrations in biological fluids. Concentrations have been assigned using R&D Systems’ Quantikine® kits. Controls are prepared in diluted porcine serum with preservatives. They contain recombinant human cytokines at low, medium and high concentrations. Controls are supplied lyophilized.
STORAGE & STABILITY
Unreconstituted Controls should be stored at 2-8 °C and are stable for at least 6 months from date of receipt. Depending on the analyte of interest, reconstituted controls may be stable when stored at < -20 °C. Users should evaluate the frozen stability of the controls in their application or discard after use.
REAGENT PREPARATION
Reconstitute each vial with the volume of deionized or distilled water indicated on the product datasheet.
PROCEDURE & EXPECTED VALUES
Controls should be assayed in the same manner as unknown specimens.
The acceptable ranges for the analytes in these controls are printed on the product datasheet. Due to possible variations in techniques and methodologies, it is recommended that each laboratory determine its own target range. Laboratories using other test systems should establish their own acceptable ranges as these assays may produce different values.
TECHNICAL HINTS & LIMITATIONS OF THE PROCEDURE
• The ranges were determined using R&D Systems’ Quantikine kits. If expected values are not obtained, verify that the lot numbers on the vials correspond with the lot numbers listed above and the correct volume of deionized or distilled water was used for reconstitution of the controls.
• The results obtained with these controls depend upon several factors associated with methods and instrumentation. Test systems other than those supplied by R&D Systems may result in values that differ from those printed on this product datasheet.
Specifications
Background: Thrombopoietin/Tpo
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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