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Sandwich ELISA analysis of Human REST matched pair antibodies
Capture: HA725193, Human REST Rabbit mAb [PSH21-38]
Detector: HA725194, Human REST Rabbit mAb [PSH21-39]
Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725193) 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 REST protein (HA211258) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725194, 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 REST in Raji extract samples based on a 1,000 µg/ml extract load.
Capture: HA725193, Human REST Rabbit mAb [PSH21-38]
Detector: HA725194, Human REST Rabbit mAb [PSH21-39]
The concentrations of REST were measured in duplicates, interpolated from the REST standard curve and corrected for sample dilution. Undiluted samples are Raji extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean REST concentration was determined to be 362.650 pg/ml in Raji extract.
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