Nuclear factor kappa light chain enhancer of activated B cells (NF-κB) p50/p65 DNA-binding assay in skeletal muscle nuclear extract
The nuclear fraction was isolated using a Nuclear Extraction kit (ab113474, Abcam) and the DNA-binding activity of p65 and p50 NF-κB subunits in the lysates was evaluated by colorimetric enzyme-linked immunosorbent assay (ELISA) using the NF-κB Transcription Factor Assay kit (ab207216, Abcam), according to the manufacturer's instructions. DNA-binding activity of combined p50/p65 was expressed as intensity based on absorbance measurement at 450 nm.