Transam enzyme linked immunosorbent assay elisa kit

Nuclear extracts were prepared from C2C12 cells or TA muscles using a Nuclear Extract Kit (Active Motif, Carlsbad, CA). NF-κB (p65) DNA-binding activity was measured using a TransAM Enzyme-Linked Immunosorbent Assay (ELISA) kit (Active Motif) according to the manufacturer’s protocol. Sample absorbance at 450 nm was measured in a spectrophotometer (Multiskan GO, Thermo Fisher Scientific). NF-κB activity is expressed as the sample absorbance at 450 nm or fold increase over the vehicle-treated controls.

NF-κB (p65) DNA-binding activity was determined using a TransAM enzyme-linked immunosorbent assay (ELISA) kit (Active Motif, Carlsbad, CA) according to the manufacturer’s protocol. In brief, nuclear extracts of C2C12 cells were prepared using the Nuclear Extract Kit (Active Motif), added to the oligonucleotide-coated plate, and incubated with the NF-κB p65-specific primary antibody (1:1000 dilution) and horseradish peroxidase-conjugated secondary antibody (1:1000 dilution) contained in the ELISA kit. After washing, the colorimetric reaction reagents were added, and sample absorbance at 450 nm was measured in a spectrophotometer (Multiskan GO, Thermo Fisher Scientific). The assay was performed in duplicate.