Natural and recombinant human Coagulation Factor III
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
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.
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.
The recovery of Coagulation Factor III spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
Citrate Plasma (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
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.
Preparation and 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).
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.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
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.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
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.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
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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