MYBBP1A Antibody / Myb-binding protein 1A (FY12713)

Immunofluorescent staining of MYBBP1A using anti-MYBBP1A antibody (green) and anti-Beta Tubulin antibody (red). MYBBP1A was detected in an immunocytochemical section of NIH/3T3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-MYBBP1A antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and Cy3 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of MYBBP1A using anti-MYBBP1A antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: rat RH35 whole cell lysates, Lane 2: mouse ovary tissue lysates, Lane 3: mouse NIH/3T3 tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MYBBP1A antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using an ECL Plus Western Blotting Substrate. A predominant doublet is observed at ~149-155 kDa, consistent with differential post-translationally modified forms of the nucleolar protein.