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Sandwich ELISA analysis of Human Growth Hormone matched pair antibodies
Capture: HA725106, Human Growth Hormone Rabbit mAb [PSH12-81]
Detector: HA725107, Human Growth Hormone Rabbit mAb [PSH12-82]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725106) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 Growth Hormone protein (HA211027) starting from 1000 pg/ml to 0 pg/ml and detect antibody (HA725107, 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 Growth Hormone in human serum samples.
Capture: HA725106, Human Growth Hormone Rabbit mAb [PSH12-81]
Detector: HA725107, Human Growth Hormone Rabbit mAb [PSH12-82]
Interpolated concentration of native Growth Hormone was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Growth Hormone concentration was determined to be 1522 pg/mL in Human serum.
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