SMC1B Antibody / Structural maintenance of chromosomes protein 1B (FY12780)

Immunofluorescent staining of SMC1B using anti-SMC1B antibody (red). SMC1B was detected in an immunocytochemical section of U2OA 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-SMC1B antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of SMC1B using anti-SMC1B antibody. Lane 1: rat testis tissue lysates, Lane 2: rat C6 whole cell lysates, Lane 3: mouse testis tissue lysates, Lane 4: mouse Neuro-2a whole cell 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-SMC1B 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 enhanced chemiluminescent. A major band is detected at ~150 kDa consistent with full-length SMC1B, with an additional band near ~250 kDa that likely represents an SDS-resistant SMC1B-containing complex or dimer enriched in testis.