Western blot using affinity purified anti-RNF25 antibody shows detection of RNF25 (arrow head) in HEK293 cells over-expressing human RNF25 (lane 1) or mouse RNF25 (lane 2). Lane 1 is a vector only control. The extracts were loaded onto a gel, followed by electrophoresis and transfer to nitrocellulose. The membrane was probed with the primary antibody diluted to 1:1,000. Personal Communication, Allan Weissman, CCR-NCI, Bethesda, MD.