All lanes : Anti-USP9x antibody ( ab19879 ) at 1 µg/ml Lane 1 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate ( ab3950 ) Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate ( ab3950 ) with Human USP9x peptide (ab20617) at 1 µg/ml Lysates/proteins at 20 µg per lane. Secondary Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/5000 dilution developed using the ECL technique Performed under reducing conditions. Predicted band size : 54, 100,105 , 290 kDa Observed band size : 100,105,290 kDa ( why is the actual band size different from the predicted? ) Additional bands at : ~50-65 kDa. We are unsure as to the identity of these extra bands.