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Sandwich ELISA analysis of Human FGF-2 matched pair antibodies.
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA721780) 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 Human FGF-2 protein (HA210834) starting from 2000 pg/ml to 0 pg/ml and detect antibody [PSH02-09]-Biotin 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 FGF-2 in Hela cell extract and U-87 MG cell extract:
The concentrations of FGF-2 were measured in duplicates, interpolated from the FGF-2 standard curve and corrected for sample dilution. Undiluted samples are Hela cell extract 0.1ug/ml and U-87 MG cell extract 0.1ug/ml. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean FGF-2 concentration was determined to be 2,106 pg/ml in Hela cell extract and 1,065 pg/ml in U-87 MG cell extract 0.1ug/ml.
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