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The binding activity of IL-1 beta (HA721279) with recombinant IL-1 beta protein.
Immobilized recombinant IL-1 beta protein at 1 μg/ml overnight at 4℃. Then blocked with 1xTBS/1%BSA for 1 hour at 37℃, and incubated with the primary antibody (HA721279) for 45min at 37℃. Then the plate was washed and incubated with 50 µl per well of Goat anti-Rabbit IgG-HRP for 0.5 hour at 37℃. 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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Standard curve of IL-1 beta matched pair antibodies:
Sandwich ELISA analysis of IL-1 beta matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody HA721278 [PS01-39] diluted in carbonate/bicarbonate buffer, at a concentration of 4 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 1% BSA/PBST blocking buffer, and incubated with serial diluted recombinant IL-1 beta protein starting from 2000 pg/ml to 31.25 pg/ml for 1 hour at 37℃. The plate was washed and incubated with 50 µl per well of detect antibody [PS00-82] (Biotin, 1:2,000) for 1 hour at 37℃. Then the plate was washed and incubated with 50 µl per well of Streptavidin-HRP for 0.5 hour at 37℃. 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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