Western blot showing detection of 0.1 µg of recombinant p27 protein. Lane 1: Molecular weight markers. Lane 2: MBP-p27 fusion protein (arrow; expected MW: 73.3 kDa). Lane 3: MBP alone. Protein was run on a 4-20% gel, then transferred to 0.45 µm nitrocellulose. After blocking with 1% BSA-TTBS overnight at 4°C, primary antibody was used at 1:1000 at room temperature for 30 min. HRP-conjugated Goat-Anti-Rabbit secondary antibody was used at 1:40,000 in MB-070 blocking buffer and imaged on the VersaDoc MP 4000 imaging system (Bio-Rad).