SYNPO2 Antibody / Synaptopodin 2 (FY12293)

Flow Cytometry analysis of human SH-SY5Y cells using anti-SYNPO2 antibody. Overlay histogram showing SH-SY5Y 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-SYNPO2 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 SYNPO2 using anti-SYNPO2 antibody. Lane 1: human SH-SY5Y 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-SYNPO2 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. Expected molecular weight of SYNPO2 ~118 kDa. In human tumor cell lysate we observe a heavy doublet at ~150 kDa. While peer-reviewed published Western blots of SYNPO2 do not consistently list a 150~160 kDa band, the gene is known to encode multiple splice variants and exhibits variable apparent migration. The upward shift likely reflects a longer isoform or post-translationally modified/complexed form of SYNPO2.