Mouse Thrombopoietin Quantikine ELISA Kit

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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Citrate Plasma (10 uL)
  • Sensitivity
    20 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Citrate Plasma)
  • Specificity
    Natural and recombinant mouse 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.
Product Summary
The Quantikine Mouse Tpo Immunoassay is a 4.5 hour solid phase ELISA designed to measure Tpo in cell culture supernates, serum, and plasma. It contains recombinant mouse Tpo and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse 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 mouse 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, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Standard Deviation9.81758.49.621.7115


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

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 97 87-115
Citrate Plasma (n=5) 104 88-113
EDTA Plasma (n=5) 104 87-116
Serum (n=5) 100 88-111
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse Tpo in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse 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
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 5 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    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.

    Showing Results 1 - 10 of 21
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    Sample Type
    1. Dynamin 2-dependent endocytosis is required for normal megakaryocyte development in mice.
      Authors: Bender M, Giannini S, Grozovsky R, Jonsson T, Christensen H, Pluthero F, Ko A, Mullally A, Kahr W, Hoffmeister K, Falet H
      Blood, 2015;125(6):1014-24.
      Species: Mouse
      Sample Type: Plasma
    2. Apoptotic effects of platelet factor VIII on megakaryopoiesis: implications for a modified human FVIII for platelet-based gene therapy.
      Authors: Greene T, Lyde R, Bailey S, Lambert M, Zhai L, Sabatino D, Camire R, Arruda V, Poncz M
      J Thromb Haemost, 2014;12(12):2102-12.
      Species: Mouse
      Sample Type: Plasma
    3. Megakaryocytes are essential for HSC quiescence through the production of thrombopoietin.
      Authors: Nakamura-Ishizu, Ayako, Takubo, Keiyo, Fujioka, Masato, Suda, Toshio
      Biochem Biophys Res Commun, 2014;454(2):353-7.
      Species: Mouse
      Sample Type: Bone Marrow Fluid
    4. Rescue of a primary myelofibrosis model by retinoid-antagonist therapy.
      Authors: Hong S, Dvorak-Ewell M, Stevens H, Barish G, Castro G, Nofsinger R, Frangos J, Shoback D, Evans R
      Proc Natl Acad Sci U S A, 2013;110(47):18820-5.
      Species: Mouse
      Sample Type: Plasma
    5. Mcl-1 and Bcl-x(L) coordinately regulate megakaryocyte survival.
      Blood, 2012;119(24):5850-8.
      Species: Mouse
      Sample Type: Serum
    6. Mcl-1 and Bcl-xL regulate Bak/Bax-dependent apoptosis of the megakaryocytic lineage at multistages.
      Cell Death Differ., 2012;19(11):1856-69.
      Species: Mouse
      Sample Type: Serum
    7. A novel role of sphingosine 1-phosphate receptor S1pr1 in mouse thrombopoiesis.
      Authors: Zhang L, Orban M, Lorenz M, Barocke V, Braun D, Urtz N, Schulz C, von Bruhl M, Tirniceriu A, Gaertner F, Proia R, Graf T, Bolz S, Montanez E, Prinz M, Muller A, von Baumgarten L, Billich A, Sixt M, Fassler R, von Andrian U, Junt T, Massberg S
      J Exp Med, 2012;209(12):2165-81.
      Species: Mouse
      Sample Type: Serum
    8. Lentiviral gene transfer regenerates hematopoietic stem cells in a mouse model for Mpl-deficient aplastic anemia.
      Authors: Heckl D, Wicke DC, Brugman MH, Meyer J, Schambach A, Busche G, Ballmaier M, Baum C, Modlich U
      Blood, 2011;117(14):3737-47.
      Species: Mouse
      Sample Type: Plasma
    9. A mouse model of hereditary folate malabsorption: deletion of the PCFT gene leads to systemic folate deficiency.
      Authors: Salojin KV, Cabrera RM, Sun W
      Blood, 2011;117(18):4895-904.
      Species: Mouse
      Sample Type: Serum
    10. Thrombopoietic effect of VPAC1 inhibition during megakaryopoiesis.
      Authors: Peeters K, Loyen S, Van Kerckhoven S, Stoffels K, Hoylaerts MF, Van Geet C, Freson K
      Br. J. Haematol., 2010;151(1):54-61.
      Species: Mouse
      Sample Type: Plasma
    11. Abnormal megakaryocyte morphology and proplatelet formation in mice with megakaryocyte-restricted MYH9 inactivation.
      Authors: Eckly A, Strassel C, Freund M, Cazenave JP, Lanza F, Gachet C, Leon C
      Blood, 2009;113(14):3182-9.
      Species: Mouse
      Sample Type: Plasma
    12. Dual requirement for the ETS transcription factors Fli-1 and Erg in hematopoietic stem cells and the megakaryocyte lineage.
      Authors: Kruse EA, Loughran SJ, Baldwin TM, Josefsson EC, Ellis S, Watson DK, Nurden P, Metcalf D, Hilton DJ, Alexander WS, Kile BT
      Proc. Natl. Acad. Sci. U.S.A., 2009;106(33):13814-9.
      Species: Mouse
      Sample Type: Serum
    13. Incomplete restoration of Mpl expression in the mpl-/- mouse produces partial correction of the stem cell-repopulating defect and paradoxical thrombocytosis.
      Authors: Lannutti BJ, Epp A, Roy J, Chen J, Josephson NC
      Blood, 2009;113(8):1778-85.
      Species: Mouse
      Sample Type: Plasma
    14. Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice.
      Authors: Xing S, Wanting TH, Zhao W, Ma J, Wang S, Xu X, Li Q, Fu X, Xu M, Zhao ZJ
      Blood, 2008;111(10):5109-17.
      Species: Mouse
      Sample Type: Plasma
    15. The lineage-c-Kit+Sca-1+ cell response to Escherichia coli bacteremia in Balb/c mice.
      Authors: Zhang P, Nelson S, Bagby GJ, Siggins R, Shellito JE, Welsh DA
      Stem Cells, 2008;26(7):1778-86.
      Species: Mouse
      Sample Type: Plasma
    16. Ratio of mutant JAK2-V617F to wild-type Jak2 determines the MPD phenotypes in transgenic mice.
      Authors: Tiedt R, Hao-Shen H, Sobas MA, Looser R, Dirnhofer S, Schwaller J, Skoda RC
      Blood, 2007;111(8):3931-40.
      Species: Mouse
      Sample Type: Plasma
    17. PECAM-1: a multifaceted regulator of megakaryocytopoiesis.
      Authors: Wu Y, Welte T, Michaud M, Madri JA
      Blood, 2007;110(3):851-9.
      Species: Mouse
      Sample Type: Serum
    18. Mutations in the cofilin partner Aip1/Wdr1 cause autoinflammatory disease and macrothrombocytopenia.
      Authors: Kile BT, Panopoulos AD, Stirzaker RA, Hacking DF, Tahtamouni LH, Willson TA, Mielke LA, Henley KJ, Zhang JG, Wicks IP, Stevenson WS, Nurden P, Watowich SS, Justice MJ
      Blood, 2007;110(7):2371-80.
      Species: Mouse
      Sample Type: Serum
    19. BclxL overexpression in megakaryocytes leads to impaired platelet fragmentation.
      Authors: Kaluzhny Y, Yu G, Sun S, Toselli PA, Nieswandt B, Jackson CW, Ravid K
      Blood, 2002;100(5):1670-8.
      Species: Mouse
      Sample Type: Serum
    20. Interleukin-6 stimulates thrombopoiesis through thrombopoietin: role in inflammatory thrombocytosis.
      Authors: Kaser A, Brandacher G, Steurer W, Kaser S, Offner FA, Zoller H, Theurl I, Widder W, Molnar C, Ludwiczek O, Atkins MB, Mier JW, Tilg H
      Blood, 2001;98(9):2720-5.
      Species: Mouse
      Sample Type: Plasma
    21. A secreted and LIF-mediated stromal cell-derived activity that promotes ex vivo expansion of human hematopoietic stem cells.
      Authors: Shih CC, Hu MC, Hu J
      Blood, 2000;95(6):1957-66.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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    Sample TestedBone marrow cells

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