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Sandwich ELISA analysis of human IFNGR1 matched pair antibodies
Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA722679) 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 standard Human IFNGR1 protein (HA210514) starting from 8,000 pg/ml to 0 pg/ml and detect antibody (HA722681B, 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 IFNGR1 in MDA-MB-231 and MCF7 extract samples based on a 1,000 µg/ml extract load.
The concentrations of IFNGR1 were measured in duplicates, interpolated from the IFNGR1 standard curve and corrected for sample dilution. Undiluted samples are MDA-MB-231 extract 50% and MCF7 extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IFNGR1 concentration was determined to be 2,358 pg/ml in MDA-MB-231 extract and 1,055 pg/ml in MCF7 extract.
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Interpolated concentrations of spiked IFNGR1 in human cell culture media samples.
The concentrations of IFNGR1 were measured in duplicates, interpolated from the IFNGR1 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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