Human Coagulation Factor III/Tissue Factor Quantikine ELISA Summary

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.

Preparation and Storage

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.

Assay Procedure

Refer to the product datasheet for the 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、Add 50 μL of Assay Diluent to each well.

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.

5、Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6、Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7、Aspirate and wash 5 times.

8、Add 100 μL Substrate Solution to each well.

9、Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.