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Sandwich ELISA analysis of Mouse IL-1 alpha matched pair antibodies
Capture: HA723526, Mouse IL-1 alpha Rabbit mAb [PSH13-40]
Detector: HA723527, Mouse IL-1 alpha Rabbit mAb [PSH13-41]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723526) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Mouse IL-1 alpha protein (HA210971) starting from 1,200 pg/ml to 0 pg/ml and detect antibody (HA723527, 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 IL-1 alpha in Raw264.7 supernatant treated or untreated with LPS for 48 hours.
Capture: HA723526, Mouse IL-1 alpha Rabbit mAb [PSH13-40]
Detector: HA723527, Mouse IL-1 alpha Rabbit mAb [PSH13-41]
Interpolated concentration of native IL-1 alpha was measured in duplicate at different sample concentrations. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1 alpha concentration was determined to be 1,546 pg/mL in neat Raw264.7 treated supernatant, undetectable in untreated Raw264.7 supernatant.
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Interpolated concentrations of spiked IL-1 alpha in cell culture media samples.
Capture: HA723526, Mouse IL-1 alpha Rabbit mAb [PSH13-40]
Detector: HA723527, Mouse IL-1 alpha Rabbit mAb [PSH13-41]
The concentrations of IL-1 alpha were measured in duplicates, interpolated from the IL-1 alpha 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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