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Sandwich ELISA analysis of Human IFN beta matched pair antibodies
Capture: HA725052, Human IFN beta Rabbit mAb [PSH17-29]
Detector: HA725053, Human IFN beta Rabbit mAb [PSH17-30]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725052) 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 IFN beta protein (HA211523) starting from 1,000 pg/ml to 0 pg/ml and detect antibody (HA725053, 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 IFN-β in A549 cell culture supernatant untreated or treated with poly I:C for 24h.
Capture: HA725052, Human IFN beta Rabbit mAb [PSH17-29]
Detector: HA725053, Human IFN beta Rabbit mAb [PSH17-30]
Interpolated concentration of native IFN-β was measured in duplicate at different sample concentrations and interpolated from the IFN-β standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IFN-β concentration was determined to be 98.9 pg/ml in A549 treated cell culture supernatant, undetectable in A549 untreated cell culture supernatant.
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