All lanes : Anti-SUZ12 antibody [SUZ220A] (ab126577) at 1/100 dilution

Lane 1 : Hek-SUZ12-V5 lysate at 30 µg
Lane 2 : Hek-mTIMP2 lysate at 30 µg
Lane 3 : Hela cell line lysate at 200 µg
Lane 4 : Jeko1 cell line lysate at 200 µg
Lane 5 : Human tonsil lysate at 200 µg
Lane 6 : Human brain lysate at 200 µg
Lane 7 : MT-2 cell line lysate at 200 µg
Lane 8 : 3T3 Mouse cell line lysate at 200 µg
Lane 9 : U266 cell line lysate at 200 µg

Predicted band size: 83 kDa
Observed band size: 83 kDa



Note: A purified version of the antibody was used at 1/100 (stock solution at 1mg/ml).

Overlay histogram showing MCF7 cells stained with ab126577 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab126577, 0.1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

All lanes : Anti-SUZ12 antibody [SUZ220A] (ab126577)

Lanes 1 & 4 : Empty vector
Lanes 2 & 5 : shSUZ12.783
Lanes 3 & 6 : shSUZ12.2076

Predicted band size: 83 kDa
Observed band size: 83 kDa



Effect of SUZ12 RNA interference (siRNA) in Jeko-1 and Z-138 mantle cell lymphoma cell lines treated with two different hairpins against SUZ12 during three days. A control cell line (empty vector) was used as negative control. Expression of SUZ12 was analyzed using ab126577. A decrease of protein expression was observed in siSUZ12 Jeko-1 and Z-138 cell extracts confirming antibody specificity. Band signals were normalized with tubulin as a loading control.

Purified version of ab126577, at 1/500, staining SUZ12 in frozen Human tonsil tissue by Immunohistochemistry.

ab126577, at a 1/30 dilution, staining SUZ12 in paraffin embedded Mouse testicle tissue by Immunohistochemistry.

Purified version of ab126577, at a 1/500 dilution, staining SUZ12 in paraffin embedded Human tonsil tissue by Immunohistochemistry.

Purified version of ab126577, at a 1/500 dilution, staining SUZ12 in paraffin embedded Human testicle tissue by Immunohistochemistry.

Purified ab126577 at 1/500 staining SUZ12 in paraffin embedded Human thymus tissue by immunohistochemistry.

Purified version of ab126577, at a 1/100 dilution, staining SUZ12 (green) in Human tonsil by Immunofluorescence. IgD antibody (red) was used at 1/50. After antigen retrieval (Tris-EDTA buffer), the slides were incubated for one hour at room temperature in a humid chamber with primary antibodies. Slides were then washed in PBS 0.5%-Tween20 3 times for 5 minutes each. The slides were incubated for 1 hour with fluorochrome-conjugated antibodies against the different Ig isotypes, diluted in PBS (dilution 1/200) in a humid chamber in the dark. Subsequently, slides were washed in PBS 0.5% Tween20 3 times for 5 minutes each. Following washing, antifading was added.

To confirm that ab126577 recognized Human SUZ12 protein, Immunohistochemistry on frozen cytospins preparations of V5-tagged Human SUZ12 expressed in Hek293T was performed. Labeling with the ab126577 confirmed the efficiency of transfection. Cytospin preparation of V5-tagged Human BTLA protein was used as a negative control.