Application
Western Blot of Rabbit anti-p35 antibody. Lane 1: p35 recombinant protein 50 ng load. Lane 2: p35 recombinant protein 100 ng load. Lane 3: p35 recombinant protein 250 ng load. Lane 4: p35 recombinant protein 500 ng load. Primary antibody: p35 antibody at 1:1,000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT. Blocking for 30 min at RT. Predicted/Observed size: 60 kDa, 60 kDa for p35.
Application
Western blot showing detection of 0.1 µg of recombinant p35 protein. Lane 1: Molecular weight markers. Lane 2: MBP-p35 fusion protein (arrow; expected MW: 69.5 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 blocking buffer and imaged on the VersaDoc MP 4000 imaging system (Bio-Rad).