SECISBP2 Antibody / Selenocysteine insertion sequence-binding protein 2 (FY13363)

Immunofluorescent staining of SECISBP2 using anti-SECISBP2 antibody (green) and anti-Beta Tubulin antibody (red). SECISBP2 was detected in immunocytochemical section of human A549 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-SECISBP2 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 SECISBP2 using anti-SECISBP2 antibody. Lane 1: human 293T whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human MCF-7 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-SECISBP2 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 dominant band at ~100 kDa is observed, consistent with the known upward size shift of SECISBP2 relative to its predicted 95 kDa mass. A second closely spaced band is present in all samples, likely representing phosphorylated or other post-translationally modified isoforms of SECISBP2. The banding pattern agrees with published reports describing SECISBP2 migration at ~100-110 kDa with multiple modified species.