Flag-tagged progerin were boiled in Laemmli buffer and the proteins were separated by 8% SDS-PAGE and blotted onto nitrocellulose. The membrane was blocked in 3% NFDM in PBS-T for 1h at RT and incubated at 4°C o/n with anti Progerin, clone 13A4D4 antibody at the indicated dilutions in 0.5% NFDM/PBS-T. After incubation with anti-mouse Fc-HRP coupled secondary antibody for 1h at RT, ECL was performed with the Western Lightning reagents. A 30sec exposure is shown.