Human Coagulation Factor III/Tissue Factor Quantikine ELISA

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DCF300
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Human Coagulation Factor III/Tissue Factor ELISA Standard Curve
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Citations (15)
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Human Coagulation Factor III/Tissue Factor Quantikine ELISA 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 (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.

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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 33.4 65.4 148 71 144 301
Standard Deviation 0.78 1.81 5 4.5 7.8 17.1
CV% 2.3 2.8 3.4 6.3 5.4 5.7

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 ELISA Linearity

Data Examples

Human Coagulation Factor III/Tissue Factor ELISA Standard Curve

Product Datasheets

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: Coagulation Factor III/Tissue Factor

Coagulation Factor III, also known as Tissue Factor (TF), Thromboplastin, and CD142, is the primary initiator of the extrinsic coagulation pathway. It is a transmembrane protein that is consitutively expressed in subendothelial cells throughout the vasculature and is inducible on endothelial cells and monocytes. Damage to the vessel wall exposes Coagulation Factor III to circulating Factor VII, triggering the proteolytic cascade that results in fibrin clot formation. Coagulation Factor III plays an important role in thrombotic diseases, disseminated intravascular coagulation, and atherosclerosis. It also plays a role in non-hemostatic inflammation, angiogenesis, and tumor progression.

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
⚠ 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. 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 for Human Coagulation Factor III/Tissue Factor Quantikine ELISA

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.

15 Citations: Showing 1 - 10
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  1. Trimethylamine N-oxide promotes tissue factor expression and activity in vascular endothelial cells: A new link between trimethylamine N-oxide and atherosclerotic thrombosis
    Authors: X Cheng, X Qiu, Y Liu, C Yuan, X Yang
    Thromb. Res., 2019;177(0):110-116.
    Species: Human
    Sample Types: Cell Lysates
  2. Alteration in endothelial permeability occurs in response to the activation of PAR2 by factor Xa but not directly by the TF-factor VIIa complex
    Authors: NE Benelhaj, A Maraveyas, S Featherby, MEW Collier, MJ Johnson, C Ettelaie
    Thromb. Res., 2019;175(0):13-20.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Upregulation of Protease-Activated Receptor 2 Promotes Proliferation and Migration of Human Vascular Smooth Muscle Cells (VSMCs)
    Authors: M Wei, Y Liu, M Zheng, L Wang, F Ma, Y Qi, G Liu
    Med. Sci. Monit., 2019;25(0):8854-8862.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Characterization of induced pluripotent stem cell-derived megakaryocyte lysates for potential regenerative applications
    Authors: A Baigger, D Eicke, Y Yuzefovych, D Pogozhykh, R Blasczyk, C Figueiredo
    J. Cell. Mol. Med., 2018;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  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, 2017;19(1):62-68.
    Species: Human
    Sample Types: Plasma
  6. Can we use biomarkers of coagulation to predict which patients with foot and ankle injury will develop deep vein thrombosis?
    Authors: BA Hickey, A Cleves, R Alikhan, N Pugh, L Nokes, A Perera
    Foot Ankle Surg, 2017;0(0):.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Capture
  7. Total circulating microparticle levels are increased in patients with deep infiltrating endometriosis
    Authors: J Munrós
    Hum. Reprod, 2016;0(0):.
    Species: Human
    Sample Types: Plasma
  8. Tissue Factor As a Predictor of Recurrent Venous Thromboembolism in Malignancy: Biomarker Analyses of the CATCH Trial
    Authors: Alok A Khorana
    J. Clin. Oncol, 2016;0(0):JCO2016674564.
    Species: Human
    Sample Types: Serum
  9. 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 Types: Cell Lysates
  10. 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 Types: Cell Culture Supernates
  11. 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 Types: Cell Lysates
  12. 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 Types: Cell Culture Supernates
  13. 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 Types: Cell Culture Supernates
  14. 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 Types: Plasma
  15. 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 Types: Cell Lysates

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Human Coagulation Factor III/Tissue Factor Quantikine ELISA
By Anonymous on 05/12/2017
Application: Sample Tested: Cell Culture Media

Selected cell lines were split into various flasks, centrifuged and supernatant used for ELISA