SCML2 Antibody / Sex comb on midleg-like protein 2 (FY13122)

Flow Cytometry analysis of human K562 cells using anti-SCML2 antibody. Overlay histogram showing K562 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-SCML2 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 (Red line) was also used as a control.

Western blot analysis of SCML2 using anti-SCML2 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human 293T 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-SCML2 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. SCML2 antibody detects a predominant band at ~85 kDa with additional weaker bands above and below in HeLa, K562, and 293T lysates. Although the calculated size is ~77 kDa, Polycomb proteins such as SCML2 often migrate larger due to post-translational modifications and chromatin-associated mobility, and alternative isoforms can resolve as closely spaced species.