Anti human hrp conjugated fc gamma specific secondary antibody

Example 2

EC50 binding of exemplary anti-IL-32 antibodies of the present invention to IL-32γ (R&D) or IL-32α (ImmunoTools), was determined by ELISA. Serial dilutions of MABs (from 100,000 ng/ml down to 1.69 ng/ml) were incubated for 2 hours with antigen-coated plates (coating overnight at 1 μg/ml in PBS, followed by wash out and blocking with 2% BSA in PBS). The plates were subsequently washed and binding of MABs was detected with anti-human HRP-conjugated Fc-gamma-specific secondary antibody (Jackson ImmunoResearch, Europe Ltd., Cambridgeshire, UK). Concentrations of MAB resulting in half of maximal binding to respective antigens (EC50, ng/ml) were calculated using Prism 4 GraphPad software on sigmoidal dose-response curves (variable slope, 4 parameters) obtained by plotting the log of the concentration versus OD 450 nm measurements; for the results see FIG. 3 and Table 4 below.

Example 2

EC50 binding of exemplary anti-IL-32 antibodies of the present invention to IL-32γ (R&D) or IL-32α (ImmunoTools), was determined by ELISA. Serial dilutions of MABs (from 100,000 ng/ml down to 1.69 ng/ml) were incubated for 2 hours with antigen-coated plates (coating overnight at 1 μg/ml in PBS, followed by wash out and blocking with 2% BSA in PBS). The plates were subsequently washed and binding of MABs was detected with anti-human HRP-conjugated Fc-gamma-specific secondary antibody (Jackson ImmunoResearch, Europe Ltd., Cambridgeshire, UK). Concentrations of MAB resulting in half of maximal binding to respective antigens (EC50, ng/ml) were calculated using Prism 4 GraphPad software on sigmoidal dose-response curves (variable slope, 4 parameters) obtained by plotting the log of the concentration versus OD 450 nm measurements; for the results see FIG. 3 and Table 4 below.