Human Coagulation Factor III/Tissue Factor Quantikine ELISA

(7 citations)
(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 (50 uL), Cell Lysates (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Citrate Plasma (50 uL), Urine (50 uL)
  • Sensitivity
    2.05 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Cell Lysates, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
  • Specificity
    Natural and recombinant human Coagulation Factor III
  • 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
QC65, Quantikine Immunoassay Control Set 804 for Human Factor III - Please Inquire
Product Summary
The Quantikine Human Coagulation Factor III/Tissue Factor Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Coagulation Factor III in cell culture supernates, cell lysates, plasma, and urine. It contains NS0-expressed recombinant human Coagulation Factor III and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Coagulation Factor III showed linear curves that were p arallel 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 Coagulation Factor III.

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, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean33.465.414871144301
Standard Deviation0.781.8154.57.817.1
CV%2.32.83.46.35.45.7

Recovery

The recovery of Coagulation Factor III spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 104 95-110
Citrate Plasma (n=4) 92 86-102
EDTA Plasma (n=4) 91 86-97
Heparin Plasma (n=4) 95 88-109
Urine (n=4) 102 96-112
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Coagulation Factor III were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Coagulation Factor III/Tissue Factor Quantikine ELISA
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Coagulation Factor III/Tissue Factor
Coagulation Factor III/Tissue Factor (TF), also known as Thromboplastin and CD142, is an integral membrane protein found in a variety of cell types. It functions as a protein cofactor/receptor of coagulation factor VII, which is synthesized in the liver and circulated in the plasma. Synthesized as a 295 amino acid precursor, TF consists of a signal peptide (residues 1 to 32) and the mature chain (residues 33 to 295), which includes a transmembrane region (residues 252 to 274) and a cytoplasmic tail (residues 275 to 295).
    • Entrez Gene IDs
      2152 (Human); 14066 (Mouse);
    • Alternate Names
      CD142 antigen; CD142; coagulation factor III (thromboplastin, tissue factor); Coagulation Factor III; F3; FLJ17960; TF; TFA; Thromboplastin; Tissue Factor;
    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. 100 µL Standard, Control, or Sample
    6.   Add 100 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:

    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.

    7 Citations: Showing 1 - 7
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    Species
    Sample Type
    1. Tissue factor is induced by interleukin-33 in human endothelial cells: a new link between coagulation and inflammation
      Sci Rep, 2016;6(0):25171.
      Species: Human
      Sample Type: Cell Lysates
    2. Loss of DGKepsilon induces endothelial cell activation and death independently of complement activation.
      Authors: Bruneau S, Neel M, Roumenina L, Frimat M, Laurent L, Fremeaux-Bacchi V, Fakhouri F
      Blood, 2015;125(6):1038-46.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Arachidonate 15-lipoxygenase enzyme products increase platelet aggregation and thrombin generation.
      Authors: Vijil C, Hermansson C, Jeppsson A, Bergstrom G, Hulten L
      PLoS ONE, 2014;9(2):e88546.
      Species: Human
      Sample Type: Cell Lysates
    4. Increased metabolite levels of glycolysis and pentose phosphate pathway in rabbit atherosclerotic arteries and hypoxic macrophage.
      Authors: Yamashita A, Zhao Y, Matsuura Y, Yamasaki K, Moriguchi-Goto S, Sugita C, Iwakiri T, Okuyama N, Koshimoto C, Kawai K, Tamaki N, Zhao S, Kuge Y, Asada Y
      PLoS ONE, 2014;9(1):e86426.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. Tissue factor-bearing exosome secretion from human mechanically stimulated bronchial epithelial cells in vitro and in vivo.
      Authors: Park J, Sharif A, Tschumperlin D, Lau L, Limbrey R, Howarth P, Drazen J
      J Allergy Clin Immunol, 2012;130(6):1375-83.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Toward engineering a human neoendothelium with circulating progenitor cells.
      Authors: Allen JB, Khan S, Lapidos KA, Ameer GA
      Stem Cells, 2010;28(2):318-28.
      Species: Human
      Sample Type: Cell Lysates
    7. Traditional risk factors and D-dimer predict incident cardiovascular disease events in chronic HIV infection.
      Authors: Ford ES, Greenwald JH, Richterman AG
      AIDS, 2010;24(10):1509-17.
      Species: Human
      Sample Type: Plasma
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 804 for Human Factor III

    Ctrl QC65
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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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     Human Coagulation Factor III/Tissue Factor Quantikine ELISA DCF300
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     Human Coagulation Factor III/Tissue Factor Quantikine ELISA DCF300
    : Human Coagulation Factor III/Tissue Factor Quantikine ELISA [DCF300]

    Summary

    Sample TestedCell Culture Media

    Other Experimental Details

    Other Experimental DetailsSelected cell lines were split into various flasks, centrifuged and supernatant used for ELISA

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