Human Thrombopoietin Quantikine ELISA Kit

(15 citations)
(2 Reviews)
  
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL)
  • Sensitivity
    18.5 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human Tpo
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC176, Quantikine Immunoassay Control Set 934 Human Thrombopoietin - Please Inquire
Product Summary
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.

Precision
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.
Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean194512983187517970
Standard Deviation17.516.526.816.339.263.9
CV%93.22.78.77.66.6

Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean20457911402035551070
Standard Deviation13.715.152.619.348.792.8
CV%6.72.64.69.58.88.7

Recovery

The recovery of Tpo spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 99 89-109
Platelet-poor EDTA Plasma (n=4) 95 85-102
Platelet-poor Heparin Plasma (n=4) 93 86-106
Serum (n=4) 102 90-111
Linearity
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.
Human Thrombopoietin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.

    • Entrez Gene IDs
      7066 (Human); 21832 (Mouse); 81811 (Rat);
    • Alternate Names
      Megakaryocyte colony-stimulating factor; Megakaryocyte growth and development factor; megakaryocyte stimulating factor; MGDF; MGDFC-mpl ligand; MK-CSF; ML; MPL ligand; MPLLGMGC163194; Myeloproliferative leukemia virus oncogene ligand; THPO; thrombopoietin nirs variant 1; Thrombopoietin; Tpo; TPOMKCSF;
    Related Research Areas
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    15 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Persistent elevated bone marrow plasma levels of thrombopoietin in patients with aplastic anemia
      Authors: Jizhou Zhang
      Cytokine, 2016;85(0):11-13.
      Species: Human
      Sample Type: Plasma
    2. Deficiency in thrombopoietin induction after liver surgery is associated with postoperative liver dysfunction.
      Authors: Haegele, Stefanie, Offensperger, Florian, Pereyra, David, Lahner, Elisabet, Assinger, Alice, Fleischmann, Edith, Gruenberger, Birgit, Gruenberger, Thomas, Brostjan, Christin, Starlinger, Patrick
      PLoS ONE, 2015;10(1):e0116985.
      Species: Human
      Sample Type: Plasma
    3. Eltrombopag for the treatment of the inherited thrombocytopenia deriving from MYH9 mutations.
      Authors: Pecci A, Gresele P, Klersy C, Savoia A, Noris P, Fierro T, Bozzi V, Mezzasoma AM, Melazzini F, Balduini CL
      Blood, 2010;116(26):5832-7.
      Species: Human
      Sample Type: Serum
    4. Thrombopoietin contributes to enhanced platelet activation in cigarette smokers.
      Authors: Lupia E, Bosco O, Goffi A
      Atherosclerosis, 2010;210(1):314-9.
      Species: Human
      Sample Type: Plasma
    5. Screening for MPL mutations in essential thrombocythemia and primary myelofibrosis: normal Mpl expression and absence of constitutive STAT3 and STAT5 activation in MPLW515L-positive platelets.
      Authors: Glembotsky AC, Korin L, Lev PR, Chazarreta CD, Marta RF, Molinas FC, Heller PG
      Eur. J. Haematol., 2010;84(5):398-405.
      Species: Human
      Sample Type: Plasma
    6. Thrombocytopenia in early malaria is associated with GP1b shedding in absence of systemic platelet activation and consumptive coagulopathy.
      Authors: de Mast Q, de Groot PG, van Heerde WL, Roestenberg M, van Velzen JF, Verbruggen B, Roest M, McCall M, Nieman AE, Westendorp J, Syafruddin D, Fijnheer R, van Dongen-Lases EC, Sauerwein RW, van der Ven AJ
      Br. J. Haematol., 2010;151(5):495-503.
      Species: Human
      Sample Type: Plasma
    7. Contributions of TRAIL-mediated megakaryocyte apoptosis to impaired megakaryocyte and platelet production in immune thrombocytopenia.
      Authors: Yang L, Wang L, Zhao CH, Zhu XJ, Hou Y, Jun P, Hou M
      Blood, 2010;116(20):4307-16.
      Species: Human
      Sample Type: Plasma
    8. Familial thrombocytosis caused by the novel germ-line mutation p.Pro106Leu in the MPL gene.
      Authors: El-Harith el-HA, Roesl C, Ballmaier M, Germeshausen M, Frye-Boukhriss H, von Neuhoff N, Becker C, Nurnberg G, Nurnberg P, Ahmed MA, Hubener J, Schmidtke J, Welte K, Stuhrmann M
      Br. J. Haematol., 2009;144(2):185-94.
      Species: Human
      Sample Type: Plasma
    9. Serum thrombopoietin compared with ZAP-70 and immunoglobulin heavy-chain gene mutation status as a predictor of time to first treatment in early chronic lymphocytic leukemia.
      Authors: Molica S, Vitelli G, Cutrona G, Todoerti K, Mirabelli R, Digiesi G, Morabito F, Neri A, Ferrarini M
      Leuk. Lymphoma, 2008;49(1):62-7.
      Species: Human
      Sample Type: Serum
    10. Helicobacter pylori eradication shifts monocyte Fcgamma receptor balance toward inhibitory FcgammaRIIB in immune thrombocytopenic purpura patients.
      Authors: Asahi A, Nishimoto T, Okazaki Y, Suzuki H, Masaoka T, Kawakami Y, Ikeda Y, Kuwana M
      J. Clin. Invest., 2008;118(8):2939-49.
      Species: Human
      Sample Type: Plasma
    11. Pharmacodynamics and Pharmacokinetics of the Novel Thrombopoietin Mimetic Peptide RWJ-800088 in Humans.
      Authors: Liem-Moolenaar M, Cerneus D, Molloy C, End D, Brown K, de Kam M, Cohen A, van Hensbergen Y, Burggraaf J
      2008;84(0):481.
      Species: Human
      Sample Type: Plasma
    12. High thrombopoietin concentrations in the cerebrospinal fluid of neonates with sepsis and intraventricular hemorrhage may contribute to brain damage.
      Authors: Reinhold A, Zhang J, Gessner R, Felderhoff-Mueser U, Obladen M, Dame C
      J. Interferon Cytokine Res., 2007;27(2):137-45.
      Species: Human
      Sample Type: CSF
    13. Circulating thrombopoietin as an in vivo growth factor for blast cells in acute myeloid leukemia.
      Authors: D&amp;amp;apos;Hondt S
      Blood, 2006;107(6):2525-30.
      Species: Human
      Sample Type: Serum
    14. Single administration of stem cell factor, FLT-3 ligand, megakaryocyte growth and development factor, and interleukin-3 in combination soon after irradiation prevents nonhuman primates from myelosuppression: long-term follow-up of hematopoiesis.
      Authors: Drouet M, Mourcin F, Grenier N, Leroux V, Denis J, Mayol JF, Thullier P, Lataillade JJ, Herodin F
      Blood, 2004;103(3):878-85.
    15. CD40-ligand stimulates myelopoiesis by regulating flt3-ligand and thrombopoietin production in bone marrow stromal cells.
      Authors: Solanilla A, Dechanet J, El Andaloussi A, Dupouy M, Godard F, Chabrol J, Charbord P, Reiffers J, Nurden AT, Weksler B, Moreau JF, Ripoche J
      Blood, 2000;95(12):3758-64.
      Species: Human
      Sample Type: Cell Culture Supernates
    Expand to show all 15 Citations
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    Average Rating: 5 (Based on 2 reviews)

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