Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit

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(1 Review)
  
  • 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), Cell Lysates (25 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Citrate Plasma (10 uL), Urine (10 uL)
  • Sensitivity
    27 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
  • Specificity
    Natural and recombinant human Thrombomodulin. This kit also recognizes Thrombomodulin complexed with Thrombin, Protein C, and Thrombin/Protein C.
  • 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
QC169, Quantikine Immunoassay Control Set 839 Human Thrombomodulin - Please Inquire
Product Summary
The Quantikine Human Thrombomodulin Immunoassay is a 4.5 hour solid phase ELISA designed to measure human Thrombomodulin in cell culture supernates, cell lysates, serum, plasma, and urine. It contains NS0-expressed recombinant human Thrombomodulin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Thrombomodulin 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 Thrombomodulin.

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, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean5711123231851810732159
Standard Deviation13.231.183.941.276.1123
CV%2.32.83.67.97.15.7

Recovery

The recovery of Thrombomodulin spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 96 88-106
Citrate Plasma (n=4) 94 85-102
EDTA Plasma (n=4) 95 86-105
Heparin Plasma (n=4) 91 85-96
Serum (n=4) 89 85-96
Urine (n=4) 99 85-115
Linearity
To assess the linearity of the assay, samples containing high concentrations of Thrombomodulin were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Thrombomodulin/BDCA-3
Thrombomodulin (TM), also known as CD141 and BDCA3, is a transmembrane expressed on vascular endothelial cells (EC), arterial smooth muscle cells, monocytes, and macrophages. It binds Thrombin and enhances the Thrombin-mediated activation of the anti-coagulant Protein C and the anti-fibrinolytic TAFI/Carboxypeptidase B2. Thrombomodulin also inhibits the ability of Thrombin to activate several pro-coagulant proteins (e.g. Fibrinogen, Factor V, Factor XIII, PAR-1). Thrombomodulin exerts multiple anti-inflammatory effects and reduces the severity of disease involving chronic inflammation. In cancer, Thrombomodulin suppresses tumor angiogenesis, growth, and epithelial-mesenchymal transition. Soluble fragments of Thrombomodulin are elevated in the serum, urine, and synovial fluid during coagulation disorders, inflammation, and organ failure.
    • Entrez Gene IDs
      7056 (Human); 21824 (Mouse); 83580 (Rat);
    • Alternate Names
      BDCA3; BDCA-3; CD141 antigen; CD141; Fetomodulin; THBD; THRM; Thrombomodulin; TM;
    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. 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.
    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.
    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. 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:

    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.

    1 Citations: Showing 1 - 1

    1. Common genetic risk factors for venous thrombosis in the chinese population.
      Am J Hum Genet, 2013;92(2):177-87.
      Species: Human
      Sample Type: Plasma
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 839 Human Thrombomodulin

    Ctrl QC169
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Cell Lysis Buffer 1 (1 x 21 mL)

    ELISA 890713 1
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    Average Rating: 5 (Based on 1 review)

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    Sample TestedCitrate Plasma

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