Human TFPI Quantikine ELISA Kit

(3 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Citrate Plasma (10 uL), Urine (50 uL)
  • Sensitivity
    6.7 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
  • Specificity
    Natural and recombinant human TFPI. This kit recognizes predominantly free TFPI and a very small percentage of the LDL- and HDL-bound TFPI.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
Product Summary
The Quantikine Human TFPI Immunoassay is a 3.5 hour solid-phase ELISA designed to measure human TFPI in cell culture supernates, plasma, and urine. It contains NS0-expressed recombinant human TFPI and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human TFPI 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 TFPI.

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, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Standard Deviation13.522.240.318.530.666.2


The recovery of TFPI spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 92 85-100
Urine (n=4) 94 85-106
To assess the linearity of the assay, samples containing or spiked with high concentrations of TFPI were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human TFPI Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: TFPI
Human TFPI is a physiological inhibitor of the extrinsic pathway of coagulation and functions in anticoagulation and anti-inflammation. It is a secreted protein with an N-terminal acidic region, three Kunitz (K) domains separated by two linker regions, and a C-terminal basic region. The first K domain inhibits coagulation factor VIIa complexed to tissue factor (TF). The second K domain inhibits factor Xa. The third K domain binds to heparin. The C-terminal basic region may have several functions. Human TFPI-2 is structurally similar to TFPI and is downregulated in several cancers.
    • Long Name
      Tissue Factor Pathway Inhibitor
    • Entrez Gene IDs
      7035 (Human); 21788 (Mouse);
    • Alternate Names
      convertin; EPI; EPITFPI1LACITFI; Extrinsic pathway inhibitor; LACI; Lipoprotein-associated coagulation inhibitor; tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor); tissue factor pathway inhibitor;
    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 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 1 hour 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.

    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.

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    Sample Type
    1. The C-terminus of tissue factor pathway inhibitor alpha is required for its interaction with factors V and Va.
      J. Thromb. Haemost., 2012;10(9):1944-6.
      Species: Human
      Sample Type: Plasma
    2. TFPIbeta is the GPI-anchored TFPI isoform on human endothelial cells and placental microsomes.
      Authors: Girard TJ, Tuley E, Broze GJ
      Blood, 2012;119(5):1256-62.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Venous thrombosis associated with gene deletion of tissue factor pathway inhibitor.
      Authors: Kentsis A, Bradwin G, Miller DT, Trenor CC
      Am. J. Hematol., 2009;84(11):775-6.
      Species: Human
      Sample Type: Plasma
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 860 for Human TFPI

    Ctrl QC167
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine Wash Buffer 1

    ELISA WA126 5
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