Human TFPI Quantikine ELISA Kit

R&D Systems | Catalog # DTFP10

R&D Systems
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Key Product Details

Assay Length

3.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL), Urine (50 µL)

Sensitivity

6.7 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
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Product Summary for Human TFPI Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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, Citrate Plasma, EDTA Plasma, Heparin 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 4.0 3.6 6.1 5.4 5.9

Recovery for Human TFPI Quantikine ELISA Kit

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 Images for Human TFPI Quantikine ELISA Kit

Human TFPI ELISA Standard Curve

Human TFPI ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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

Alternate Names

EPI, LACI

Entrez Gene IDs

7035 (Human); 21788 (Mouse)

Gene Symbol

TFPI

Additional TFPI Products

Product Documents for Human TFPI Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human TFPI Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human TFPI Quantikine ELISA Kit

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Protocols

View specific protocols for Human TFPI Quantikine ELISA Kit (DTFP10):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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