Human TFPI Quantikine ELISA Kit Summary

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.

Preparation and Storage

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.

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.