ab111893 labelling Smac / Diablo (green) in the cytoplasm of A431 cells by Immunocytochemistry/Immunofluorescence. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4 ºC. A DyLight-conjugated anti-mouse IgG was used as the secondary antibody. Red (phalloidiin) - F-actin, Blue - nuclei. Images were taken at a magnification of 60x.

ab111893 labelling Smac / Diablo (green) in the cytoplasm of MCF-7 cells by Immunocytochemistry/Immunofluorescence. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4 ºC. A DyLight-conjugated anti-mouse IgG was used as the secondary antibody. Red (phalloidiin) - F-actin, Blue - nuclei. Images were taken at a magnification of 60x.

ab111893 labelling Smac / Diablo (green) in the cytoplasm of PC12 cells by Immunocytochemistry/Immunofluorescence. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody (1:100 in 3% BSA-PBS) overnight at 4 ºC. A DyLight-conjugated anti-mouse IgG was used as the secondary antibody. Red (phalloidiin) - F-actin, Blue - nuclei. Images were taken at a magnification of 60x.

All lanes : Anti-Smac/Diablo antibody [SMAC 17 1-87] (ab111893) at 1/1000 dilution

Lane 1 : 293 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : PC12 cell lysate

Lysates/proteins at 25 µg per lane.

Predicted band size: 27 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?