Human Thrombospondin-1 Quantikine ELISA Kit

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DTSP10
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Thrombospondin-1 ELISA Kit ELISA
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Human Thrombospondin-1 Quantikine ELISA Kit Summary

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 (25 uL), Serum (10 uL), Platelet-poor EDTA Plasma (25 uL), Platelet-poor Heparin Plasma (25 uL), Saliva (50 uL), Human Milk (50 uL)
Sensitivity
0.944 ng/mL
Assay Range
7.8 - 500 ng/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Human Milk)
Specificity
Natural and recombinant human Thrombospondin-1
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 Human Thrombospondin-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Thrombospondin-1 in cell culture supernates, serum, plasma, saliva, and human milk. It contains NS0-expressed recombinant human Thrombospondin-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Thrombospondin-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Thrombospondin-1.

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, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 71.9 142 282 75.8 151 294
Standard Deviation 4.34 9.56 18 4.71 8.51 17.6
CV% 6 6.7 6.4 6.2 5.6 6

Recovery

The recovery of Thrombospondin-1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 95 86-101
Platelet-poor EDTA Plasma (n=4) 94 89-99
Platelet-poor Heparin Plasma (n=4) 93 87-104

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Thrombospondin-1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Thrombospondin-1 ELISA Linearity

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Thrombospondin-1

Thrombospondins (TSP) are secreted multidomain glycoproteins with many putative functions including modulating cell adhesion, proliferation, migration, and angiogenesis. At least five TSPs exist: TSP-1, -2, -3, -4, and cartilage oligomeric glycoprotein (COMP)/TSP-5. The family can be divided into two subgroups. The first, TSP-1 and -2, are homotrimeric molecules, each monomer containing an N-terminal/heparin-binding domain, a coiled-coil oligomerization domain, a von Willebrand factor-type/procollagen homology domain, three type 1 thrombospondin repeats (TSR), three EGF-like repeats, seven TSP type 3 repeats, and a C-terminal globular domain. TSP-1 and -2 are thought to mediate interactions between cells and the extracellular matrix, and as such are matricellular proteins. In contrast, TSP-3, -4, and COMP/TSP-5 are homopentamers that lack the N-terminal structure of TSP-1 and -2, but have a similar arrangement of EGF and TSP type 3 repeats, and the C-terminal domain characteristic of the TSP family.

Entrez Gene IDs:
7057 (Human); 21825 (Mouse)
Alternate Names:
THBS; THBS1; THBS-1; thrombospondin 1; Thrombospondin1; Thrombospondin-1; thrombospondin-1p180; TSP-1; TSP1thrombospondin-1; TSPTSP-1
⚠ 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.

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. 100 µL Assay Diluent
  4.   Add 100 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 on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Thrombospondin-1 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.

26 Citations: Showing 1 - 10
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  1. Elevated ADAMTS13 Activity is Associated with Poor Postoperative Outcome in Patients Undergoing Liver Resection
    Authors: S Haegele, J Fuxsteiner, D Pereyra, C Koeditz, B Rumpf, C Schuetz, C Schwarz, C Brostjan, T Gruenberge, P Starlinger
    Sci Rep, 2018;8(1):16823.
    Species: Human
    Sample Types: Plasma
  2. MicroRNA-206 suppresses TGF-? signalling to limit tumor growth and metastasis in lung adenocarcinoma
    Authors: K Watt, D Newsted, E Voorand, RJ Gooding, A Majewski, P Truesdell, B Yao, T Tuschl, N Renwick, AW Craig
    Cell. Signal., 2018;50(0):25-36.
    Species: Human
    Sample Types: Tissue Homogenates
  3. Optimized plasma preparation is essential to monitor platelet-stored molecules in humans
    Authors: M Mussbacher, WC Schrottmai, M Salzmann, C Brostjan, JA Schmid, P Starlinger, A Assinger
    PLoS ONE, 2017;12(12):e0188921.
    Species: Human
    Sample Types: Plasma
  4. Vitamin D supplementation lowers thrombospondin-1 levels and blood pressure in healthy adults
    Authors: AT Amaraseker, B Assadi-Kha, S Liu, M Black, G Dymmott, NM Rogers, AL Sverdlov, JD Horowitz, DTM Ngo
    PLoS ONE, 2017;12(5):e0174435.
    Species: Human
    Sample Types: Plasma
  5. Hedgehog signalling pathway orchestrates angiogenesis in triple-negative breast cancers
    Authors: C Di Mauro, R Rosa, V D'Amato, P Ciciola, A Servetto, R Marciano, RC Orsini, L Formisano, S De Falco, V Cicatiello, M Di Bonito, M Cantile, F Collina, A Chambery, BM Veneziani, S De Placido, R Bianco
    Br. J. Cancer, 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Inflammatory Differences in Plaque Erosion and Rupture in Patients With ST-Segment Elevation Myocardial Infarction
    Authors: S Chandran, J Watkins, A Abdul-Aziz, M Shafat, PA Calvert, KM Bowles, MD Flather, SA Rushworth, AD Ryding
    J Am Heart Assoc, 2017;6(5):.
    Species: Human
    Sample Types: Plasma
  7. Aureobasidium pullulans produced ?-glucan is effective to enhance Kurosengoku soybean extract induced Thrombospondin-1 expression
    Authors: D Muramatsu, M Okabe, A Takaoka, H Kida, A Iwai
    Sci Rep, 2017;7(1):2831.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Unaltered Angiogenesis-Regulating Activities of Platelets in Mild Type 2 Diabetes Mellitus despite a Marked Platelet Hyperreactivity
    PLoS ONE, 2016;11(9):e0162405.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Combining bevacizumab and chemoradiation in rectal cancer. Translational results of the AXEBeam trial.
    Authors: Verstraete M, Debucquoy A, Dekervel J, Van Pelt J, Verslype C, Devos E, Chiritescu G, Dumon K, D'Hoore A, Gevaert O, Sagaert X, Van Cutsem E, Haustermans K
    Br J Cancer, 2015;112(8):1314-25.
    Species: Human
    Sample Types: Whole Blood
  10. Net platelet angiogenic activity (NPAA) correlates with progression and prognosis of non-small cell lung cancer.
    Authors: Yao L, Dong H, Luo Y, Du J, Hu W
    PLoS ONE, 2014;9(4):e96206.
    Species: Human
    Sample Types: Cell Lysates
  11. BMP4/Thrombospondin-1 loop paracrinically inhibits tumor angiogenesis and suppresses the growth of solid tumors.
    Authors: Tsuchida, R, Osawa, T, Wang, F, Nishii, R, Das, B, Tsuchida, S, Muramatsu, M, Takahashi, T, Inoue, T, Wada, Y, Minami, T, Yuasa, Y, Shibuya, M
    Oncogene, 2014;33(29):3803-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Thrombospondin-1 production is enhanced by Porphyromonas gingivalis lipopolysaccharide in THP-1 cells.
    Authors: Gokyu M, Kobayashi H, Nanbara H, Sudo T, Ikeda Y, Suda T, Izumi Y
    PLoS ONE, 2014;9(12):e115107.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Long-term survival and biomarker correlates of tasquinimod efficacy in a multicenter randomized study of men with minimally symptomatic metastatic castration-resistant prostate cancer.
    Authors: Armstrong A, Haggman M, Stadler W, Gingrich J, Assikis V, Polikoff J, Damber J, Belkoff L, Nordle O, Forsberg G, Carducci M, Pili R
    Clin Cancer Res, 2013;19(24):6891-901.
    Species: Human
    Sample Types: Plasma
  14. Thrombospondin-1 expression in melanoma is blocked by methylation and targeted reversal by 5-Aza-deoxycytidine suppresses angiogenesis.
    Authors: Lindner D, Wu Y, Haney R, Jacobs B, Fruehauf J, Tuthill R, Borden E
    Matrix Biol, 2013;32(2):123-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. MicroRNA-16 affects key functions of human endothelial progenitor cells.
    Authors: Goretti E, Rolland-Turner M, Leonard F, Zhang L, Wagner D, Devaux Y
    J Leukoc Biol, 2013;93(5):645-55.
  16. Predictors of thromboxane levels in patients with non-ST-elevation acute coronary syndromes on chronic aspirin therapy.
    Thromb. Haemost., 2012;108(1):133-9.
    Species: Human
    Sample Types: Plasma
  17. Metronomic activity of CD44-targeted hyaluronic acid-paclitaxel in ovarian carcinoma.
    Clin. Cancer Res., 2012;18(15):4114-21.
    Species: Complex Species Category
    Sample Types: Plasma
  18. Pharmacodynamic and pharmacogenetic angiogenesis-related markers of first-line FOLFOXIRI plus bevacizumab schedule in metastatic colorectal cancer.
    Authors: Loupakis F, Cremolini C, Fioravanti A
    Br. J. Cancer, 2011;104(8):1262-9.
    Species: Human
    Sample Types: Plasma
  19. Thrombospondin-1 triggers cell migration and development of advanced prostate tumors.
    Authors: Firlej V, Mathieu JR, Gilbert C, Lemonnier L, Nakhle J, Gallou-Kabani C, Guarmit B, Morin A, Prevarskaya N, Delongchamps NB, Cabon F
    Cancer Res., 2011;71(24):7649-58.
    Species: Human
    Sample Types: Cell Lysates
  20. Escherichia coli K1 promotes the ligation of CD47 with thrombospondin-1 to prevent the maturation of dendritic cells in the pathogenesis of neonatal meningitis.
    Authors: Mittal R, Gonzalez-Gomez I, Prasadarao NV
    J. Immunol., 2010;185(5):2998-3006.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Platelet-derived growth factor (PDGF)-PDGF receptor interaction activates bone marrow-derived mesenchymal stromal cells derived from chronic lymphocytic leukemia: implications for an angiogenic switch.
    Authors: Ding W, Knox TR, Tschumper RC
    Blood, 2010;116(16):2984-93.
    Species: Human
    Sample Types: Platelet-Poor Plasma
  22. Tasquinimod (ABR-215050), a quinoline-3-carboxamide anti-angiogenic agent, modulates the expression of thrombospondin-1 in human prostate tumors.
    Authors: Olsson A, Bjork A, Vallon-Christersson J, Isaacs JT, Leanderson T
    Mol. Cancer, 2010;9(0):107.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Thrombospondin-1: a unique marker to identify in vitro platelet activation when monitoring in vivo processes.
    Authors: Starlinger P, Moll HP
    J. Thromb. Haemost., 2010;0(0):.
    Species: Human
    Sample Types: Serum
  24. Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
    Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
    PLoS ONE, 2009;4(11):e7916.
    Species: Human
    Sample Types: Plasma
  25. Estrogen regulation of thrombospondin-1 in human breast cancer cells.
    Int. J. Cancer, 2009;125(0):1045.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Cytokine secretion by human mesenchymal stem cells cocultured with damaged corneal epithelial cells.
    Authors: Oh JY, Kim MK, Shin MS, Wee WR, Lee JH
    Cytokine, 2009;46(1):100-3.
    Species: Human
    Sample Types: Cell Culture Supernates

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