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Sandwich ELISA analysis of Human Myeloperoxidase matched pair antibodies
Capture: HA723673, Human Myeloperoxidase Rabbit mAb [PSH14-78]
Detector: HA723674, Human Myeloperoxidase Rabbit mAb [PSH14-79]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723673) 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 Human Myeloperoxidase protein (HA211022) starting from 12,000 pg/ml to 0 pg/ml and detect antibody (HA723674, 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 Myeloperoxidase in human samples.
Capture: HA723673, Human Myeloperoxidase Rabbit mAb [PSH14-78]
Detector: HA723674, Human Myeloperoxidase Rabbit mAb [PSH14-79]
Interpolated concentration of native Myeloperoxidase was measured in duplicate at different sample concentrations and interpolated from the Myeloperoxidase standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Myeloperoxidase concentration was determined to be 432 ng/mL in HL-60 cell supernatant. There was no detectable signal in Hela cell supernatant.
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Interpolated concentrations of spiked Myeloperoxidase in cell culture media samples.
Capture: HA723673, Human Myeloperoxidase Rabbit mAb [PSH14-78]
Detector: HA723674, Human Myeloperoxidase Rabbit mAb [PSH14-79]
The concentrations of Myeloperoxidase were measured in duplicates, interpolated from the Myeloperoxidase 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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