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Sandwich ELISA analysis of Mouse Factor XII matched pair antibodies
Capture: HA723443, Mouse/Rat Factor XII Rabbit mAb [PSH12-21]
Detector: HA723444, Mouse/Rat Factor XII Rabbit mAb [PSH12-23]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723443) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 Mouse Factor XII protein (HA210984) starting from 18,000 pg/ml to 0 pg/ml and detect antibody (HA723444, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 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 Factor XII in mouse samples.
Capture: HA723443, Mouse/Rat Factor XII Rabbit mAb [PSH12-21]
Detector: HA723444, Mouse/Rat Factor XII Rabbit mAb [PSH12-23]
Interpolated concentration of native Factor XII was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Factor XII concentration was determined to be 22,555,940 pg/mL in mouse serum and 20,935 pg/ml in mouse liver Supernatant.
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Interpolated concentrations of spiked Factor XII in cell culture media samples.
Capture: HA723443, Mouse/Rat Factor XII Rabbit mAb [PSH12-21]
Detector: HA723444, Mouse/Rat Factor XII Rabbit mAb [PSH12-23]
The concentrations of Factor XII were measured in duplicates, interpolated from the Factor XII standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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Interpolated concentrations of native Factor XII in rat samples.
Capture: HA723443, Mouse/Rat Factor XII Rabbit mAb [PSH12-21]
Detector: HA723444, Mouse/Rat Factor XII Rabbit mAb [PSH12-23]
Interpolated concentration of native Factor XII was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Factor XII concentration was determined to be 16,655,930 pg/mL in rat serum.
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