Western Blot of Rabbit Anti-NF-Y(A subunit) Antibody. Lane 1: CHO-7 cells in the absence (I) of cholesterol. Lane 2: CHO-7 cells in the presence (S) of cholesterol. Load: Equivalent aliquots of chromatin from each sample containing ~50 μg of total protein per lane. Primary antibody: anti-NF-YA antibody at 1:1000 for overnight at 4ºC. Secondary antibody: HRP conjugated Goat-anti-Rabbit IgG [H&L] secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: doublet of ~ 42 kDa for NF-Y A.