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Human Thrombopoietin Quantikine ELISA Kit

Catalog #: DTP00B
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Human Thrombopoietin/Tpo ELISA Serum/Plasma Standard Curve
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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Thrombopoietin Quantikine ELISA Kit Summary

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.

Sample Values

Serum/Platelet-poor Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human Tpo in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean of Detectable (pg/mL)% DetectableRange (pg/mL)
Serum (n=38)74.274ND-228
Pletelet-poor EDTA plasma (n=34)91.821ND-196
Platelet-poor Heparin plasma (n=37)82.819ND-168
ND=Non-detectable

Cell Culture Supernates:
Human peripheral blood cells (1 x 106 cells/mL) were cultured in DMEM supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and assayed for levels of human Tpo. No detectable levels were observed.

HepG2 human hepatocellular carcinoma cells were cultured in DMEM supplemented with 10% fetal bovine serum. An aliquot of the culture supernate was removed, assayed for human Tpo, and measured 81.9 pg/mL.

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

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 204 579 1140 203 555 1070
Standard Deviation 13.7 15.1 52.6 19.3 48.7 92.8
CV% 6.7 2.6 4.6 9.5 8.8 8.7

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 194 512 983 187 517 970
Standard Deviation 17.5 16.5 26.8 16.3 39.2 63.9
CV% 9 3.2 2.7 8.7 7.6 6.6

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/Tpo ELISA Linearity

Scientific Data

N/A Human Thrombopoietin/Tpo ELISA Serum/Plasma Standard Curve View Larger

Human Thrombopoietin/Tpo ELISA Serum/Plasma Standard Curve

N/A Human Thrombopoietin/Tpo ELISA Cell Culture Supernate Standard Curve View Larger

Human Thrombopoietin/Tpo ELISA Cell Culture Supernate Standard Curve

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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; MKCSF; MK-CSF; ML; MPL ligand; MPLLG; MPLLGMGC163194; Myeloproliferative leukemia virus oncogene ligand; THCYT1; THPO; thrombopoietin nirs variant 1; Thrombopoietin; Tpo; TPOMKCSF
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human Thrombopoietin Quantikine ELISA Kit

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.

32 Citations: Showing 1 - 10
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  1. Novel modes of MPL activation in triple-negative myeloproliferative neoplasms
    Authors: SE Samaraweer, T Geukens, DA Casolari, T Nguyen, C Sun, S Bailey, S Moore, J Feng, AW Schreiber, WT Parker, AL Brown, C Butcher, PG Bardy, M Osborn, HS Scott, D Talaulikar, CS Grove, CN Hahn, RJ D'Andrea, DM Ross
    Pathology, 2022-08-05;0(0):.
    Species: Human
    Sample Types: Serum
  2. Cell-autonomous megakaryopoiesis associated with polyclonal hematopoiesis in triple-negative essential thrombocythemia
    Authors: T Inano, M Araki, S Morishita, M Imai, Y Kihara, M Okuda, Y Yang, M Ito, S Osaga, H Mano, Y Edahiro, T Ochiai, K Misawa, Y Fukuda, J Ando, N Komatsu
    Scientific Reports, 2021-09-06;11(1):17702.
    Species: Human
    Sample Types: Serum
  3. Can Coagulation System Disorders and Cytokine and Inflammatory Marker Levels Predict the Temporary Clinical Deterioration or Improvement of Septic Patients on ICU Admission?
    Authors: GA Lavranou, S Mentzelopo, P Katsaounou, I Siempos, I Kalomenidi, A Geranaki, C Routsi, S Zakynthino
    Journal of Clinical Medicine, 2021-04-07;10(8):.
    Species: Human
    Sample Types: Serum
  4. Histopathological Chromogranin A-Positivity Is Associated with Right-Sided Colorectal Cancers and Worse Prognosis
    Authors: Z Herold, M Dank, M Herold, P Nagy, K Rosta, A Somogyi
    Cancers, 2020-12-29;13(1):.
    Species: Human
    Sample Types: Plasma
  5. A Pilot Study on the Effect of Anti-Thrombopoietin Antibody on Platelet Count in Patients with Type 2 Diabetes
    Authors: T Fukuda, M Hamaguchi, T Osaka, Y Hashimoto, E Ushigome, M Asano, M Yamazaki, E Fukuda, K Yamaguchi, K Ogawa, N Goshima, M Fukui
    Molecules, 2020-04-04;25(7):.
    Species: Human
    Sample Types: Serum
  6. Multilevel defects in the hematopoietic niche in essential thrombocythemia
    Authors: T Sun, M Ju, X Dai, H Dong, W Gu, Y Gao, R Fu, X Liu, Y Huang, W Liu, Y Chi, W Wang, H Li, Y Zhou, L Shi, R Yang, L Zhang
    Haematologica, 2019-07-09;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Eltrombopag for the treatment of inherited thrombocytopenias: a phase 2 clinical trial
    Authors: C Zaninetti, P Gresele, A Bertomoro, C Klersy, E De Candia, D Veneri, S Barozzi, T Fierro, MA Alberelli, V Musella, P Noris, F Fabris, CL Balduini, A Pecci
    Haematologica, 2019-07-04;0(0):.
    Species: Human
    Sample Types: Serum
  8. Host Immunological Effects of Partial Splenic Embolization in Patients with Liver Cirrhosis
    Authors: Y Matsukiyo, H Nagai, T Matsui, Y Igarashi
    J Immunol Res, 2018-07-15;2018(0):1746391.
    Species: Human
    Sample Types: Serum
  9. MECOM-associated syndrome: a heterogeneous inherited bone marrow failure syndrome with amegakaryocytic thrombocytopenia
    Authors: M Germeshaus, P Ancliff, J Estrada, M Metzler, E Ponstingl, H Rütschle, D Schwabe, RH Scott, S Unal, A Wawer, B Zeller, M Ballmaier
    Blood Adv, 2018-03-27;2(6):586-596.
    Species: Human
    Sample Types: Plasma
  10. Thrombopoietin levels in Quebec platelet disorder-Implications for the mechanism of thrombocytopenia
    Authors: CPM Hayward, S Tasneem, GE Rivard
    Int J Lab Hematol, 2018-02-01;0(0):.
    Species: Human
    Sample Types: Plasma
  11. Thrombopoietin and its receptor expression in pediatric patients with chronic immune thrombocytopenia
    Authors: S Li, J Shao, M Xia, N Zhang, J Yang, H Li, H Jiang
    Hematology, 2018-01-09;0(0):1-6.
    Species: Human
    Sample Types: Plasma
  12. Acquired amegakaryocytic thrombocytopenia previously diagnosed as idiopathic thrombocytopenic purpura in a patient with hepatitis C virus infection
    Authors: S Ichimata, M Kobayashi, K Honda, S Shibata, A Matsumoto, H Kanno
    World J. Gastroenterol., 2017-09-21;23(35):6540-6545.
    Species: Human
    Sample Types: Serum
  13. Bone marrow failure unresponsive to bone marrow transplant is caused by mutations in THPO
    Authors: A Seo, M Ben-Harosh, M Sirin, J Stein, O Dgany, J Kaplelushn, M Hoenig, U Pannicke, M Lorenz, K Schwarz, C Stockklaus, T Walsh, S Gulsuner, MK Lee, A Sendamarai, M Sanchez-Bo, MC King, H Cario, AE Kulozik, KM Debatin, A Schulz, H Tamary, A Shimamura
    Blood, 2017-05-30;0(0):.
    Species: Human
    Sample Types: Serum
  14. A new form of inherited thrombocytopenia due to monoallelic loss of function mutation in the thrombopoietin gene
    Authors: P Noris, C Marconi, D De Rocco, F Melazzini, T Pippucci, G Loffredo, T Giangregor, A Pecci, M Seri, A Savoia
    Br. J. Haematol., 2017-05-03;0(0):.
    Species: Human
    Sample Types: Serum
  15. Alteration Analysis of Bone Marrow Mesenchymal Stromal Cells from De Novo Acute Myeloid Leukemia Patients at Diagnosis
    Authors: L Desbourdes, J Javary, T Charbonnie, N Ishac, J Bourgeais, A Iltis, JC Chomel, A Turhan, F Guilloton, K Tarte, MV Demattei, E Ducrocq, F Rouleux-Bo, E Gyan, O Hérault, J Domenech
    Stem Cells Dev., 2017-04-10;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Thrombopoietin Secretion by Human Ovarian Cancer Cells
    Authors: S Besbes, S Shah, I Al-Dybiat, S Mirshahi, H Helfer, H Najah, C Fourgeaud, M Pocard, I Ghedira, J Soria, M Mirshahi
    Int J Cell Biol, 2017-03-30;2017(0):1873834.
    Species: Human
    Sample Types: Cell Lysates
  17. Elevated thrombopoietin and platelet indices confirm active thrombopoiesis but fail to predict clinical severity of puumala hantavirus infection
    Authors: Outi Laine
    Medicine (Baltimore), 2016-12-01;95(52):e5689.
    Species: Human
    Sample Types: Serum
  18. Persistent elevated bone marrow plasma levels of thrombopoietin in patients with aplastic anemia
    Authors: Jizhou Zhang
    Cytokine, 2016-06-03;85(0):11-13.
    Species: Human
    Sample Types: Plasma
  19. 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, 2015-01-22;10(1):e0116985.
    Species: Human
    Sample Types: Plasma
  20. 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., 2010-10-19;151(5):495-503.
    Species: Human
    Sample Types: Plasma
  21. 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, 2010-09-15;116(26):5832-7.
    Species: Human
    Sample Types: Serum
  22. 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, 2010-07-29;116(20):4307-16.
    Species: Human
    Sample Types: Plasma
  23. 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., 2010-01-22;84(5):398-405.
    Species: Human
    Sample Types: Plasma
  24. Thrombopoietin contributes to enhanced platelet activation in cigarette smokers.
    Authors: Lupia E, Bosco O, Goffi A
    Atherosclerosis, 2009-11-20;210(1):314-9.
    Species: Human
    Sample Types: Plasma
  25. 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., 2008-11-19;144(2):185-94.
    Species: Human
    Sample Types: Plasma
  26. 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
    2008-10-01;84(0):481.
    Species: Human
    Sample Types: Plasma
  27. 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., 2008-08-01;118(8):2939-49.
    Species: Human
    Sample Types: Plasma
  28. 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, 2008-01-01;49(1):62-7.
    Species: Human
    Sample Types: Serum
  29. 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., 2007-02-01;27(2):137-45.
    Species: Human
    Sample Types: CSF
  30. Circulating thrombopoietin as an in vivo growth factor for blast cells in acute myeloid leukemia.
    Authors: D&amp;apos;Hondt S
    Blood, 2005-11-29;107(6):2525-30.
    Species: Human
    Sample Types: Serum
  31. 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, 2003-10-02;103(3):878-85.
    Species: Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque)
    Sample Types: Plasma
  32. 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, 2000-06-15;95(12):3758-64.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human Thrombopoietin Quantikine ELISA Kit
By Ann Cullinane on 09/05/2017
Sample Tested: Platelet-poor Citrate Plasma
Human Thrombopoietin Quantikine ELISA Kit
By Jakub Zmajkovic on 07/20/2017
Sample Tested: Cell Culture Supernates
Human Thrombopoietin Quantikine ELISA Kit DTP00B