Application
Western blot using GeneTex's anti-NAG-1 monoclonal antibody. The blot shows detection of recombinant NAG-1 protein present in Pichia pastoris whole cell lysates: lane 1 - yeast cell lysate expressing NAG-1 H variant with SUMO expression tag at 36 kDa; lane 2 - yeast cell lysate expressing NAG-1 D variant with SUMO expression tag at 36 kDa; lane 3 - yeast cell lysate expressing NAG-1 H variant; and lane 4 - yeast cell lysate expressing NAG-1 D variant. Recombinant NAG-1 proteins without SUMO correspond to monomer (15 kDa) and dimer (30 kDa) bands as indicated by the arrowheads. All lysates were run under reducing conditions. Primary antibody was used at a 1:1,000 dilution in TBS containg 1% BSA and 0.2% Tween, and reacted overnight at 4°C. For detection, a 1:40,000 dilution of peroxidase conjugated Goat-anti-Mouse IgG secondary antibody for 30 min at room temperature. Molecular weight estimation was made by comparison to prestained MW markers.