Human TFPI Quantikine ELISA Kit

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DTFP10
Control Products Available
Human TFPI ELISA Standard Curve
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Product Details
Procedure
Citations (10)
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Human TFPI Quantikine ELISA Kit Summary

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.

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.

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, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 299 554 1108 301 569 1127
Standard Deviation 13.5 22.2 40.3 18.5 30.6 66.2
CV% 4.5 4 3.6 6.1 5.4 5.9

Recovery

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

Linearity

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 ELISA Linearity

Scientific Data

Human TFPI ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TFPI

Tissue Factor Pathway Inhibitor (TFPI) is a key regulator of the extrinsic coagulation pathway. It inhibits coagulation through the formation of a quaternary complex with Coagulation Factor X, Coagulation Factor III/Tissue Factor, and Coagulation Factor VII, thereby preventing the activation of Factor X. TFPI also exhibits anti-angiogenic and anti-metastatic effects. It is produced by vascular endothelial cells and circulates as a free molecule, on platelet surfaces, and in complex with LDL and HDL particles. Abnormal TFPI concentrations in the circulation are associated with thrombosis, disseminated intravascular coagulation, and sepsis. TFPI can enhance anti-thrombotic treatment in sepsis, inflammatory diseases, and cardiovascular diseases.

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; TFPI; tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor); tissue factor pathway inhibitor
⚠ 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 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.

Citations for Human TFPI 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.

10 Citations: Showing 1 - 10
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  1. Different Profiles of Cytokines, Chemokines and Coagulation Mediators Associated with Severity in Brazilian Patients Infected with Dengue Virus
    Authors: VE Fiestas So, NR da Costa F, CF Dos Santos, G Corrêa, MDC Cipitelli, M Dornelas R, LJ de Souza, PV Damasco, RV da Cunha, FB Dos Santos, LM de Oliveir, EL de Azeredo
    Viruses, 2021-09-08;13(9):.
    Species: Human
    Sample Types: Plasma
  2. Postprandial factor VII activation does not increase plasma concentrations of prothrombin fragment 1�+�2 in patients with morbid obesity
    Authors: LE Landgrebe, CB Juhl, V Andersen, L Moitinho-S, C Bang, EM Bladbjerg
    Thromb. Res., 2020-09-01;196(0):260-267.
    Species: Human
    Sample Types: Serum
  3. Testosterone treatment and association with thrombin generation and coagulation inhibition in Klinefelter syndrome: A cross-sectional study
    Authors: S Chang, D Biltoft, A Skakkebæk, J Fedder, A Bojesen, MV Bor, CH Gravholt, AB Münster
    Thromb. Res., 2019-08-19;182(0):175-181.
    Species: Human
    Sample Types: Plasma
  4. Dual inhibition of thrombin and activated factor X attenuates disseminated intravascular coagulation and protects organ function in a baboon model of severe Gram-negative sepsis
    Authors: H Schöchl, M van Griens, S Heitmeier, V Laux, U Kipman, J Roodt, S Bahrami, H Redl
    Crit Care, 2017-03-13;21(1):51.
    Species: Primate - Papio ursinus (Chacma baboon)
    Sample Types: Plasma
  5. Increased circulating procoagulant and anticoagulant factors as TF and TFPI according to severity or infecting serotypes in human dengue infection
    Authors: E Leal de Az, VE Solórzano, DB de Oliveir, CF Marinho, LJ de Souza, RV da Cunha, PV Damasco, CF Kubelka, LM de-Oliveir
    Microbes Infect, 2016-08-31;19(1):62-68.
    Species: Human
    Sample Types: Plasma
  6. MiR-205 and MiR-373 Are Associated with Aggressive Human Mucinous Colorectal Cancer
    Authors: Annette Eyking
    PLoS ONE, 2016-06-06;11(6):e0156871.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Tissue factor is induced by interleukin-33 in human endothelial cells: a new link between coagulation and inflammation
    Sci Rep, 2016-05-04;6(0):25171.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. The C-terminus of tissue factor pathway inhibitor alpha is required for its interaction with factors V and Va.
    J. Thromb. Haemost., 2012-09-01;10(9):1944-6.
    Species: Human
    Sample Types: Plasma
  9. TFPIbeta is the GPI-anchored TFPI isoform on human endothelial cells and placental microsomes.
    Authors: Girard TJ, Tuley E, Broze GJ
    Blood, 2011-12-05;119(5):1256-62.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Venous thrombosis associated with gene deletion of tissue factor pathway inhibitor.
    Authors: Kentsis A, Bradwin G, Miller DT, Trenor CC
    Am. J. Hematol., 2009-11-01;84(11):775-6.
    Species: Human
    Sample Types: Plasma

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