All lanes : Anti-Piccolo antibody - Synaptic Marker ( ab20664 ) at 1 µg/ml Lane 1 : Brain (Rat) Tissue Lysate Lane 2 : Brain (Mouse) Tissue Lysate Lane 3 : Brain (Rat) Tissue Lysate with Rat Piccolo peptide (ab23400) at 1 µg/ml Lane 4 : Brain (Mouse) Tissue Lysate with Rat Piccolo peptide (ab23400) at 1 µg/ml Lysates/proteins at 30 µg per lane. Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Performed under reducing conditions. Predicted band size : 520 kDa Observed band size : 275 kDa ( why is the actual band size different from the predicted? ) Additional bands at : 31 kDa,60 kDa. We are unsure as to the identity of these extra bands. Exposure time : 4 minutes