Quantifying Nuclear NF-κB Transcription Factors

Activated transcription factors NF-κBp65 and NF-κBp50 in nuclear extracts of transfected or non-transfected MDA-MB-231 cells were determined using a highly sensitive Transcription Factor Assay Kit (catalog # ab133128, Abcam, England). Transcription Factor NF-κBp65 combo positive control (Abcam) and Transcription Factor NF-κBp50 combo positive control (Abcam) were also used as experimental controls. The plates with the final reaction were read at 450 nm using an automatic Zenythmicroplate reader (Anthos 3100, Salzburg, Austria). The nuclear extract of transfected or non-transfected MDA-MB-231 cells was prepared as described previously (37 (link), 38 (link)).