Fig. 2

A standard curve with rHfIL-1β by dehydration coating method. To establish an ELISA system for HfIL-1β, first a standard curve was established with rHfIL-1β using the dehydration coating method. The purified rHfIL-1β was diluted with the coating buffer with the following final concentrations: 0, 3, 6, 9, 12, 15, 15, 18, 21, and 24 ng/mL. Fifty-microliter of serially diluted rHfIL-1β were added to each well of Nunc MaxiSorp® flat-bottom 96-well plate, followed by incubation at 60°C for 2 h. Then, the plate was sequentially incubated with anti-ChIL-1β pAb (1:1000) and goat anti-rabbit antibody (1: 2000). The HRP signal was developed with TMB solution for 30 min. The coating buffer itself was used as negative control. Values represent the mean of three independent experiments. Error bars represent standard error of the mean. The dashed line indicates the threshold line, representing the value of negative control and limitation of the developed ELISA system (OD450 = 0.038)