Anti-Oct-2 antibody [EPR542] (ab92458) at 1/1000 dilution + Daudi (Human Burkitt's lymphoma cell line) lysate at 10 µg

Secondary
HRP conjugated goat anti-rabbit antibody at 1/2000 dilution

Predicted band size: 51 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?

ab92458 at a 1:250 dilution staining Oct-2 in human tonsil (paraffin embedded section) by immunohistochemistry.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

Immunocytochemistry analysis of Raji (human Burkitt's lymphoma B lymphocyte) labeling 37530 with purified ab92458 at 1/500 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.19 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control: PBS instead of the primary antibody.

Overlay histogram showing Raji (Human Burkitt's lymphoma cell line) cells stained with ab92458 (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 (ab92458, 1:50) 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) (1 µg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events was performed.