All lanes : Anti-HtrA2 / Omi antibody [EPR22] (ab75982) at 1/5000 dilution

Lane 1 : Jurkat cell lysate
Lane 2 : 293T cell lysate
Lane 3 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 49 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?
Additional bands at: 43 kDa. We are unsure as to the identity of these extra bands.

This data was developed using ab75982, the same antibody clone in a different buffer formulation.

Secondary antibody - goat anti-rabbit HRP (ab6721).

This data was developed using ab75982, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human colon using ab75982 at 1/100-1/250 dilution. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

This data was developed using ab75982, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with ab75982 (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 (ab75982, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor® 488 (IgG H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.