Rabbit anti human histone h3 polyclonal antibody

Proteins were quantified using the bicinchoninic acid method. Then, 50 µg protein/lane was subjected to electrophoresis in a 5% stacking gel and 12% separation gel and were then transferred onto a PVDF membrane. The membrane was blocked in 5% non-fat milk at room temperature for 2 h and then treated with rabbit anti-human PML polyclonal antibody (cat. no. ab53773; 1:1,000), rabbit anti-human NE polyclonal antibody (cat. no. ab68672; 1:1,000; both Abcam, Cambridge, MA, USA), rabbit anti-human Histone H3 polyclonal antibody (cat. no. P30266; 1:5,000; Abmart, Shanghai, China) and mouse anti-human β-actin antibodies (cat. no. BM0627; 1:4,000; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) in 5% non-fat milk at 4°C overnight. Following washing in TBST twice (10 min for each) and in TBS for 10 min, the membrane was treated with goat anti-rabbit (cat. no. ZB-2301) or goat anti-mouse (cat. no. ZB-2305) IgG horseradish peroxidase (HRP)-linked secondary antibodies (Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.; 1:1,000, in 5% non-fat milk) at 37°C for 1 h. The membrane was then washed again as described above. Visualization was performed as described previously (10 (link),11 (link)).