Electrophoretic mobility shift assay (EMSA) for NF-κB DNA-binding

An electrophoretic mobility shift assay (EMSA) kit was used to measure the NF-κB DNA-binding activity. Extracted DNA was incubated with a p65 biotin end-labeled NF-κB oligonucleotide probe (forward: 5′-AGT TGA GGG GAC TTT CCC AGG C-3′; reverse: 5′-G CCT GGG AAA GTC CCC TCA ACT-3′) in binding buffer for 15 min at room temperature. Then, the p65 biotin-labeled oligonucleotide complex was incubated for a further 15 min. Gel electrophoresis was used for the separation of the sample in Tris buffer solution and ECL was used to detect the bands.