Mouse Thrombopoietin Quantikine ELISA Kit

  (27 citations)
(2 Reviews)
    
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Assay Procedure
Citations (27)
FAQs
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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.

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, Serum, EDTA Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 123 455 2364 119 457 2389
Standard Deviation 9.8 17 58.4 9.6 21.7 115
CV% 8 3.7 2.5 8.1 4.7 4.8

Recovery

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
Linearity
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.
 Thrombopoietin/Tpo [HRP]
Product Datasheets

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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.
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.

27 Citations: Showing 1 - 10
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Species
Sample Type
  1. D-dencichine Regulates Thrombopoiesis by Promoting Megakaryocyte Adhesion, Migration and Proplatelet Formation
    Authors: S Ding, M Wang, S Fang, H Xu, H Fan, Y Tian, Y Zhai, S Lu, X Qi, F Wei, G Sun, X Sun
    Front Pharmacol, 2018;9(0):297.
    Species: Mouse
    Sample Type: Plasma
  2. The thrombopoietin/MPL axis is activated in the Gata1(low) mouse model of myelofibrosis and is associated with a defective RPS14 signature
    Authors: M Zingariell, L Sancillo, F Martelli, F Ciaffoni, M Marra, L Varricchio, RA Rana, C Zhao, JD Crispino, AR Migliaccio
    Blood Cancer J, 2017;7(6):e572.
    Species: Mouse
    Sample Type: Plasma
  3. Repercussion of Megakaryocyte-Specific Gata1 Loss on Megakaryopoiesis and the Hematopoietic Precursor Compartment
    PLoS ONE, 2016;11(5):e0154342.
    Species: Mouse
    Sample Type: Plasma
  4. 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
  5. 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
  6. Erythropoietin guides multipotent hematopoietic progenitor cells toward an erythroid fate.
    Authors: Grover, Amit, Mancini, Elena, Moore, Susan, Mead, Adam J, Atkinson, Deborah, Rasmussen, Kasper D, O'Carroll, Donal, Jacobsen, Sten Eir, Nerlov, Claus
    J Exp Med, 2014;211(2):181-8.
    Species: Mouse
    Sample Type: Serum
  7. Rasa3 controls megakaryocyte Rap1 activation, integrin signaling and differentiation into proplatelet.
    Authors: Molina-Ortiz P, Polizzi S, Ramery E, Gayral S, Delierneux C, Oury C, Iwashita S, Schurmans S
    PLoS Genet, 2014;10(6):e1004420.
  8. Platelet production proceeds independently of the intrinsic and extrinsic apoptosis pathways.
    Authors: Josefsson, Emma C, Burnett, Deborah, Lebois, Marion, Debrincat, Marlyse, White, Michael, Henley, Katya J, Lane, Rachael, Moujalled, Diane, Preston, Simon P, O'Reilly, Lorraine, Pellegrini, Marc, Metcalf, Donald, Strasser, Andreas, Kile, Benjamin
    Nat Commun, 2014;5(0):3455.
    Species: Mouse
    Sample Type: Serum
  9. 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
  10. 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
  11. 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
  12. Mcl-1 and Bcl-x(L) coordinately regulate megakaryocyte survival.
    Blood, 2012;119(24):5850-8.
    Species: Mouse
    Sample Type: Serum
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
  21. 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
  22. 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
  23. 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
  24. 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
  25. 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
  26. 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
  27. 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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 Mouse Thrombopoietin Quantikine ELISA Kit MTP00 Array MTP00
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  Anonymous 08/17/2017
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 Mouse Thrombopoietin Quantikine ELISA Kit MTP00
: Mouse Thrombopoietin Quantikine ELISA Kit [MTP00].

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Sample Tested Serum and Plasma
 Mouse Thrombopoietin Quantikine ELISA Kit MTP00 Array MTP00
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Very Good
  Anonymous 07/12/2016
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 Mouse Thrombopoietin Quantikine ELISA Kit MTP00
: Mouse Thrombopoietin Quantikine ELISA Kit [MTP00].

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Sample Tested Bone marrow cells

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