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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.
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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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