SEC61B Antibody / Sec61 subunit beta (FY12360)

Flow Cytometry analysis of PC-3 cells using anti-SEC61B antibody. Overlay histogram showing PC-3 cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SEC61B antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of SEC61B using anti-SEC61B antibody. Lane 1: human whole cell lysates, Lane 2: human PC-3 whole cell lysates, Lane 3: human 293T whole cell lysates, Lane 4: human Hela whole cell lysates, Lane 5: rat C6 whole cell lysates, Lane 6: mouse C2C12 whole cell lysates, Lane 7: mouse RAW264.7 whole cell lysates, Lane 8: mouse 3T3-L1 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-SEC61B antibody at 0.25 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. SEC61B (~10–11 kDa predicted) was detected as a band at ~14 kDa, likely reflecting its small size combined with membrane integration and potential post-translational modification or complex retention which slows migration in SDS–PAGE.