All lanes : Anti-HRPT2 antibody ( ab20661 ) at 1 µg/ml Lane 1 : 20ug HeLa cell lysate Lane 2 : 20ug HEK293 lysate Lane 3 : 20ug HeLa cell lysate with Human HRPT2 peptide (ab23385) at 2 µg/ml Lane 4 : 20ug HEK293 cell lysate with Human HRPT2 peptide (ab23385) at 2 µg/ml Secondary Alexa Fluor goat polyclonal to rabbit IgG (700) at 1/10000 dilution Observed band size : 60 kDa ( why is the actual band size different from the predicted? ) Additional bands at : 100 kDa (possible cross reactivity, but this band is not blocked),34 kDa (possible cleavage fragment,cross reactivity),52 kDa (possible cross reactivity, but this band is not blocked).