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Sandwich ELISA analysis of Human Lumican matched pair antibodies
Capture: HA725317, Human Lumican Rabbit mAb [PSH19-04]
Detector: HA725318, Human Lumican Rabbit mAb [PSH19-05]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725317) 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 Lumican protein (HA211171) starting from 80,000pg/ml to 0 pg/ml and detect antibody (HA725318, Biotin, 0.05 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 Lumican in human serum samples.
Capture: HA725317, Human Lumican Rabbit mAb [PSH19-04]
Detector: HA725318, Human Lumican Rabbit mAb [PSH19-05]
The concentrations of Lumican were measured in duplicates, interpolated from the Lumican standard curve and corrected for sample dilution. Undiluted samples are human serum 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Lumican concentration was determined to be 172,905 pg/ml in human serum.
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