All lanes : Anti-MiTF antibody ( ab20663 ) at 1 µg/ml Lane 1 : HeLa whole cell lysate Lane 2 : HeLa nuclear lysate Lane 3 : A431 whole cell lysate Lane 4 : HeLa whole cell lysate with Human MiTF peptide (ab21454) at 1 µg/ml Lane 5 : HeLa nuclear lysate with Human MiTF peptide (ab21454) at 1 µg/ml Lane 6 : A431 whole cell lysate with Human MiTF peptide (ab21454) at 1 µg/ml Lysates/proteins at 1 µg/ml per lane. Secondary Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution Observed band size : 52 kDa ( why is the actual band size different from the predicted? ) Additional bands at : 25 kDa,37 kDa. We are unsure as to the identity of these extra bands.