Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH22-45] to Human Tissue Factor antibody (Capture) (HA725401) and Recombinant Human Tissue Factor protein (HA211432) as the standard. The reference range value is 3.9-500 pg/mL.
ELISA
Use at an assay dependent concentration.
Target
Function
Tissue factor, also called platelet tissue factor or Coagulation factor III,[5] is a protein present in subendothelial tissue and leukocytes which plays a major role in coagulation and, in humans, is encoded by F3 gene. Its role in the blood clotting is the initiation of thrombin formation from the zymogen prothrombin. Thromboplastin defines the cascade that leads to the activation of factor X—the tissue factor pathway. In doing so, it has replaced the previously named extrinsic pathway in order to eliminate ambiguity. The F3 gene encodes tissue factor also known as coagulation factor III, which is a cell surface glycoprotein. This factor enables cells to initiate the blood coagulation cascades, and it functions as the high-affinity receptor for the coagulation factor VII. The resulting complex provides a catalytic event that is responsible for initiation of the coagulation protease cascades by specific limited proteolysis. Unlike the other cofactors of these protease cascades, which circulate as nonfunctional precursors, this factor is a potent initiator that is fully functional when expressed on cell surfaces. There are three distinct domains of this factor: extracellular, transmembrane, and cytoplasmic. This protein is the only one in the coagulation pathway for which a congenital deficiency has not been described. In addition to the membrane-bound tissue factor, soluble form of tissue factor was also found which results from alternatively spliced tissue factor mRNA transcripts, in which exon 5 is absent and exon 4 is spliced directly to exon 6.
Background References
1. Jeon HM et al. Tissue factor is a critical regulator of radiation therapy-induced glioblastoma remodeling. Cancer Cell. 2023 Aug
2. Subramaniam S et al. Tissue factor in COVID-19-associated coagulopathy. Thromb Res. 2022 Dec
Sandwich ELISA analysis of Human Tissue Factor matched pair antibodies
Capture: HA725401, Human Tissue Factor Rabbit mAb [PSH22-45] Detector: HA725402, Human Tissue Factor Rabbit mAb [PSH22-46]
Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725401) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human Tissue Factor protein (HA211432) starting from 500 pg/ml to 0 pg/ml and detect antibody (HA725402, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Interpolated concentrations of native Tissue Factor in Hep G2 and 3T3-L1 lys samples based on a 1,000 µg/ml extract load.
Capture: HA725401, Human Tissue Factor Rabbit mAb [PSH22-45] Detector: HA725402, Human Tissue Factor Rabbit mAb [PSH22-46]
The concentrations of Tissue Factor were measured in duplicates, interpolated from the Tissue Factor standard curve and corrected for sample dilution. Undiluted samples are Hep G2 lys 0.5mg/ml and 3T3-L1 lys 1mg/ml. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Tissue Factor concentration was determined to be 170 pg/ml in Hep G2 lys and undetectable in 3T3-L1 lys.
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