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Sandwich ELISA analysis of Human NPTX2 matched pair antibodies
Capture: HA725187, Human NPTX2 Rabbit mAb [PSH21-45]
Detector: HA725188, Human NPTX2 Rabbit mAb [PSH21-46]
Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725187) 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 NPTX2 protein (HA211175) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA725188, 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 NPTX2 in human serum samples.
Capture: HA725187, Human NPTX2 Rabbit mAb [PSH21-45]
Detector: HA725188, Human NPTX2 Rabbit mAb [PSH21-46]
The concentrations of NPTX2 measured in duplicate and interpolated from the NPTX2 standard curve and corrected for sample dilution. Undiluted samples are human serum 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean NPTX2 concentration was determined to be 12,890 pg/ml in human serum.
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