Western blot using Mdm2 polyclonal antibody (Cat # PAB10274) is shown to detect a band (arrow) corresponding to mouse Mdm2 protein present in mouse MEF cells (Lane 2), but not human kidney HEK293 cells (lane1). Approximately 35 ug of lysate was separated by 4-20% Tris Glycine SDS-PAGE. After blocking the membrane with 5% normal goat serum, 0.5% BLOTTO in PBS, the membrane was probed for overnight at 4° with the primary antibody diluted to 1:500 in 1% normal goat serum, 0.1% BLOTTO in PBS. The membrane was washed and reacted with a 1 : 10,000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] for 45 min at roomtemperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers indicated at the right (700 nm channel, red). IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc.