Anti ahr bml sa210

MEF cells were scraped in 300-μl 2× sample buffer (125 mM Tris HCl, 4% SDS, 16% glycerol, 10% β-mercaptoethanol, 0.002% bromophenol blue) and boiled for 5 min. Total protein concentrations were measured by Bio-Rad assay following the manufacturer’s instructions. For Western blotting, equal amounts of protein from each sample, up to 30 μg per lane, were separated on precast Tris-Glycine gels (Invitrogen) and transferred to nitrocellulose membranes. Primary antibodies and their dilutions were as follows: anti-AHR (BML-SA210; Enzo Life Sciences), 1:1000; anti-ARNT (sc-17811, Santa Cruz Biotechnology), 1:200; anti-CUL4B (12916-1-AP; ProteinTech), 1:1000; anti-β actin (A5441, Sigma) 1:50,000; anti-histone-3, H3 (H0164, Sigma), 1:30,000, and anti-α/β tubulin (2168, Cell signaling), 1:1000. Secondary antibodies were peroxidase-conjugated goat anti-rabbit (A6154, Sigma) or mouse IgG kappa binding protein (sc-516102S, Santa Cruz Biotechnology). Protein bands were detected with ECL Western Blotting Detection reagents (GE Healthcare). Band intensities were measured by densitometry and were normalized to β-actin, α/β tubulin, or histone 3 (H3) levels using GeneTools analysis software (Syngene).