Anti-MST3 antibody [EP1468Y] (ab51137) at 1/5000 dilution + HeLa cell lysate at 10 µg

Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution

Predicted band size: 49 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?

Ab51137 (1:100) staining human MST3 in human brain tissue by immunohistochemistry using paraffin embedded tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunofluorescent staining of HeLa cells using ab51137 (1:100).

Overlay histogram showing HeLa cells stained with ab51137 (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 (ab51137, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.