Anti-MDM2 Antibody - Western Blot. Western blot using affinity purified Anti-MDM2 (Rabbit) 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 (lane 1). 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 1:10000 dilution of IRDye800 conjugated Gt-a-Rabbit IgG [H&L] ( for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers indicated at the right (700 nm channel, red). IRDye800 fluorescence image was captured using the Ddyssey Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.