Rabbit anti human creb1

HCT116 cell layers on glass coverslips were fixed for 15 min using 4% paraformaldehyde, permeabilized for 20 min in PBS containing 0.2% Triton X-100, and then blocked for 2 h with PBS containing 1% BSA and 0.5% goat serum at 37°C. The cells were incubated with primary antibody at 4°C overnight. The antibodies mouse anti-human RRM2, rabbit anti-human CREB1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). After rinsing with PBS, and then probed with FITC- or TRITC- conjugated secondary antibodies (Santa Cruz, CA, USA) for 1 h at 37°C. The nuclei were stained with DAPI (Sigma) for 15 min. The slides were mounted and visualized by a fluorescence microscope (AX70, Olympus, Tokyo, Japan). The images are representative of triplicate independent experiments.

The whole cell lysate were analyzed with the antibodies mouse anti-human RRM2, rabbit anti-human CREB1, rabbit anti-human E2F1, mouse anti-human Flag, mouse anti-human cmyc, mouse anti-human GAPDH (Santa Cruz Biotechnology), IRDye® 800CW- or IRDye® 680-conjugated secondary antibodies (LI-COR, Lincoln,NE) were used for staining and then detected by an Odyssey® infrared imaging system (LI-COR).