RELA activity in the over-expression lysate (LY402895) and a vector-transfected control lysate was measured in a colorimetric DNA-binding assay. Double-stranded oligonucleotide containing the RELA consensus DNA-binding sequence was incubated with dilutions of the over-expression lysate and RELA bound to the oligo was captured onto the surface of a microtiter plate. After washing, bound RELA was detected with an anti-RELA primary antibody followed by an HRP-labeled secondary antibody. After initial color development, the reaction was quenched and the color intensity was measured at 450nm. The data show high levels of RELA DNA binding activity in the over-expression lysate, but almost no DNA-binding activity in the control lysate. Overexpression cell lysates are prepared from HEK293T cells transfected with RC220780 using transfection reagent MegaTran 2.0 (Cat# TT210002).