Anti Bmi1

Immunofluorescence using Rockland's affinity purified goat anti Bmi1 shows nuclear staining (green) of methanol fixed (100%, 5 min) HepG2 cells. The cells were blocked and permeabilized in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS Tween for 1h prior to incubation with the primary antibody (1:200 dilution) overnight at +4C and detected with a 488nm fluorescent dye conjugated secondary Ab. Cell nuclei are stained with DAPI (blue) and plasma membranes are stained with WGA (red).

Western blot using Rockland's Affinity Purified anti-Bmi1 antibody shows detection of a band ~37 kDa corresponding to human Bmi1 (arrowhead).？？ Approximately 20 ug of a U2OS whole cell lysate (bone osteosarcoma) was separated by 4-20% SDS-PAGE and transferred onto nitrocellulose.？ After blocking in PBS containing 5% nonfat dry milk, the membrane was probed overnight at 4 C with the primary antibody diluted to 1:1,000 in PBS containing 1% nonfat dry milk.？ The membrane was washed and reacted with a 1:20,000 dilution of IRDyeTM800 conjugated Rb-a-Goat IgG [H&L] MX (605-432-013) for 45 min at room temperature. IRDyeTM800 fluorescence image was captured using the OdysseyR Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc.？ Other detection systems will yield similar results.