Figure 5

ELISA Analysis of Immune Sera for DEK Reactivity. 96-well plates were coated with recombinant DEK or EGFP (10 μg/ml) produced in bacterial vectors. Wells were blocked, and 100 μl of test serum (as described on the x-axis), diluted 1:100, was added to each well. Sera used for this assay was the same serum used for cDNA library screening, and was pooled from 5 immunized mice. Serum from naïve A/J mice, mice treated with PC61 alone, or AGN2a-CD80/CD86 vaccine alone was derived from parallel experimental groups used in the immunization protocol. Following incubation at 37°C for 1 hour, secondary rabbit anti-mouse IgG (H+L) was used to detect bound antibody, as detailed in Methods. Also included was anti-human DEK monoclonal antibody (Anti-DEK-MAb) used at 0.5 μg/ml. Error bars show standard deviations for triplicate sample wells in the same assay.